Hemostatic compositions and uses therefor

ABSTRACT

The present invention relates generally to the field of hemostasis, including methods, compositions, and devices that can be employed to achieve hemostasis at an increased rate. More specifically the present invention relates to hemostatic compositions that achieve a hemostatic effect at a distance from the site of application of the composition, and a method for administering such a composition to effectively reduce localized vascular complications associated with treating a breach or puncture in a vein or artery and reduce the time to achieve hemostasis.

This application is a divisional of U.S. patent application Ser. No.10/749,330, filed on Dec. 31, 2003, now U.S. Pat. No. 7,931,637 whichclaims priority of U.S. provisional Patent Application No. 60/437,349,filed on Dec. 31, 2002, each of which is incorporated herein byreference in its entirety.

1. FIELD OF THE INVENTION

The present invention relates generally to the field of hemostasis,including methods, compositions, and devices that can be employed toachieve hemostasis at an increased rate or in a reduced period of time.More specifically the present invention relates to hemostaticcompositions that achieve a hemostatic effect at a distance from thesite of application of the composition, and a method for administeringsuch a composition to effectively reduce localized vascularcomplications associated with treating a breach or puncture in a vein orartery. The hemostatic compositions can comprise one or morevasoconstrictors or one or more coagulants, or a combination of one ormore vasoconstrictors with one or more coagulants. The hemostaticcompositions can also comprise a compound that has both vasoconstrictiveproperties and coagulant properties. The hemostatic compositions can beapplied in conjunction with a barrier-forming material to moreeffectively achieve hemostasis and reduce the occurrence of localizedvascular complications.

2. BACKGROUND

Localized vascular complications associated with catheterization of avein or artery are common side effects with many medical procedures.Such complications including hemorrhaging of blood vessels, delayedhemostasis time, hematoma, pseudoaneurysm, and arteriovenous (AV)fistula formation can be life threatening. Since cardiac catheterizationremains the primary technique for diagnosing coronary artery disease andis used to help delineate coronary anatomy, the information gained isoften a medical necessity, despite the risks. Traditionally, theapplication of pressure to a vein or artery effected by a breach orpuncture was the preferred method for reducing complications, howevercompression of a vein or artery after cardiac catheterization can bevery painful for the patients. Compression bandages with weights may beapplied for 4-8 hours following hemostasis and patients can beimmobilized up to 24 hours. Patients frequently develop back pain andurinary retention in addition to the localized vascular complications.

Two approaches have recently been used to reduce localized vascularcomplications, namely percutaneous closure/sealant devices andhemostatic patches.

Numerous hemostatic patches have been invented to treat superficialwounds on skin or on the surface of internal organs. These patches areeffective at the site where the patch contacts the skin or contacts thesurface of an internal organ. Application of the patches in treatment ofcardiac catheterization presents a problem in that the underlyingpuncture in the artery or vein is not directly effected by the activecomponents of the patches. Thus, the vein or artery puncture maycontinue to bleed after the skin puncture wound is sealed and thepossibility of hematoma formation or other localized vascularcomplications exists. U.S. Pat. No. 6,056,970 discloses a hemostaticbiocompatable composition and a method of application wherein thecomposition is topically maintained in contact with the wound on theskin surface, preferably with light pressure, for a period of time forclotting to occur at the interface between the composition and thewound. A related composition is disclosed in U.S. Pat. No. 6,361,551,where a wound-contacting fabric accelerates clot formation at aninterface between a wound surface and the hemostatic fabric. Thehemostatic fabric is composed of collagen fibers and can comprisenumerous hemostatic agents. U.S. Pat. No. 5,564,849 also discloses asimilar patch that accelerates clot formation at an interface between askin or organ wound surface and the hemostatic patch. The patchcomprises active amounts of thrombin and epsilon aminocaproic acid(EACA).

Several compositions that can act as hemostatic agents and typicallyinclude collagen or fibrin are known (Falstrom et al., 1997,Catheterization and Cardiovasular Diagnosis 41:79-84; Hoekstra et al.,1998, Biomaterials. 19:1467-1471; Prior et al., 2000, Journal ofBiomedical Materials Research. 53(3):252-257). The main disadvantage ofsuch compositions is their limited use in treating breaches or puncturesin veins or arteries at a distance. In order to treat breaches orpunctures in veins or arteries at a distance beneath the skin surfacewith such compositions, it is necessary to apply the compositions in aninvasive manner so that the compositions are in contact with a breach orpuncture in a vein or artery. Since such compositions are effective atthe site of contact, topical treatment at a breach or puncture in a veinor artery that is found at some distance from the skin surface is notpossible. U.S. Pat. No. 4,394,373 discloses compositions that act ascoagulants and may be used to promote clotting of a wound by placing thecompositions in contact with the wound where the composition comprisesliquid or powder chitosan. U.S. Pat. No. 5,510,102 disclosescompositions that act as coagulants and may be used to promote clottingof a wound by placing the compositions in contact with the wound wherethe composition comprises platelet rich plasma plus a biocompatablepolymer that is a hemostatic agent such as alginate. The compositions ofU.S. Pat. Nos. 4,394,373 and 5,510,102 are either applied directly tothe wound surface, in the case of treatment of a superficial wound or inthe case of a puncture in an artery left by a catheterization procedure,the compositions are typically injected into the soft tissue surroundingthe arterial puncture site, so that the composition is in contact withthe puncture site. The injection of the compositions is an invasiveprocedure that can lead to complications such as swelling, furtherdamage to blood vessels and tissue, and increased risk of infection.

Hemostatic patches are effective at the interface of the wound and thepatch. Thus, in the case of a puncture in a vein or artery caused bycatheterization, when a patch causes clotting at the skin surface woundsite, the deeper internal puncture in the vein or artery continues tohemorrhage increasing the likelihood that a hematoma or other vascularcomplications will occur. Hemostatic patches have not been demonstratedto be effective at a distance in treating punctures resulting fromcatheterization, but nonetheless hemostatic patches have numeroussurgical applications where the patches or fabrics can be applieddirectly to the wound surfaces on organs exposed during surgicalprocedures.

Another approach to reducing localized vascular complications associatedwith catheterization procedures has been the development of devices toaid in the application of hemostatic compositions percutaneously, sothat a hemostatic composition is administered in contact with thepuncture site. Kipshidze et al. (U.S. Pat. No. 5,437,292) and Van Tasselet al. (U.S. Pat. No. 6,193,670 B1) each invented a device for sealingcatheter puncture sites in blood vessels. Such a device typicallyadministers a plug composed of collagen or fibrin that extends from thesite of the puncture in the vein or artery through the catheter sheathtract to the skin surface. Numerous variations of such device have beendeveloped where the puncture site is sealed with a suture (Wood et al.U.S. Pat. No. 5,746,755), or where the puncture in the blood vessel issealed from within the blood vessel lumen (Redmond et al. U.S.2002/0006429 A1). Gwechenberger, Silber, Camenzind, and Sanborn eachprovided a comparison of sealant devices employed with compressioncompared with manual compression alone (Camenzind et al, 1994, Journalof the American College of Cardiology. 24(3):655-662; Gwechenberger etal., 1997, Angiology. 48(2):121-126; Sanborn et al., 1993, Journal ofthe American College of Cardiology. 22(5):1273-1279; Silber et al.,1998, Catheterization and Cardiovasular Diagnosis 43:421-427). Nearly,all commercially available devices and patches are suggested for use inconjunction with compression. The main disadvantage of such devices isthe invasive manner in which the compositions or sutures are applied.

Although several hemostatic devices are commercially available,complication rates with theses closure devices are similar toconventional manual compression (Olade et al., 2002, Emedicine Journal,3(3):1-13). Devices and methods which do not suture the catheterarterial puncture, but plug the catheter tract with collagen from theskin surface to the arterial puncture site have also been shown to be nomore effective than manual compression alone (Camenzind et al., 1994,Journal of the American College of Cardiology. 24(3):655-662). Whetherarterial puncture closure devices or hemostatic patches are used totreat a breach or puncture in a vein or artery, there remains a need forreducing the incidence of life threatening complications associated withcatheterization and other procedures which has been a persistent medicalproblem which bespeaks the need for new treatments.

3. SUMMARY OF THE INVENTION

The present invention provides methods and compositions for treatmentfor a breach or puncture in a vein or artery that would decrease theoccurrence of hematomas and other localized vascular complications.Without being bound by any theory, the inventors believe that thecompositions and methods of the invention are effective in tissuesbeyond the site of contact with the skin surface wound relative to thedevices and patches of the art. Thus, the compositions and methods ofthe invention provide the advantage and decreased likelihood oflocalized vascular complications. The main feature of the methods andcompositions of the invention over the existing hemostatic patches anddevices is the non-invasive manner in which the compositions functionand can be applied.

Accordingly, the present invention provides a method for treating abreach or puncture in a vein or artery of a patient that comprises: a)applying topically to the patient's skin over a wound contiguous withthe breach or puncture in the vein or artery a composition comprising aneffective amount of a vasoconstrictor, wherein the vasoconstrictor doesnot comprise a poly-β-1→4 N-acetylglucosamine polymer or derivativethereof; and concurrently b) applying compression to the breached orpunctured vein or artery, wherein a cessation or reduction of blood flowout of the breach or puncture in the vein or artery is achieved at agreater rate or in a reduced period of time than applying compression inconjunction with a topical barrier-forming material without avasoconstrictor. In related embodiments, the composition furthercomprises a coagulant.

In other embodiments of the present invention, a method for achieving acessation of blood flow or sealing of a breach or puncture in a vein orartery and a cessation of blood flow or sealing of a skin surface woundthat is contiguous with the breach or puncture comprises: a) applyingtopically to the patient's skin over a wound contiguous with a breach orpuncture in a vein or artery a composition comprising a vasoconstrictoror coagulant, wherein the vasoconstrictor or coagulant does not comprisea poly-β-1→4 N-acetylglucosamine polymer or derivative thereof; b)concurrently applying compression to the breached or punctured vein orartery; and c) recording the amount of blood flow from the wound and thepuncture, wherein an amount of the vasoconstrictor or coagulant iseffective to increase sealing or increase cessation of blood flow fromthe breach or puncture in the vein or artery and increase sealing orincrease cessation of blood flow from the skin surface wound, incomparison to applying compression in conjunction with a topicalbarrier-forming material without a vasoconstrictor.

In other embodiments of the present invention, a method for treating abreach or puncture in a vein or artery of a patient, comprises: a)applying topically to the patient's skin over a wound contiguous withthe breach or puncture in the vein or artery a composition comprising aneffective amount of a coagulant, wherein the coagulant does not comprisea poly-β-1→4 N-acetylglucosamine polymer or derivative thereof; andconcurrently b) applying compression to the breached or punctured veinor artery, wherein a cessation or reduction of blood flow out of thebreach or puncture in the vein or artery is achieved at a greater rateor in a reduced period of time than applying compression in conjunctionwith a topical barrier-forming material without a coagulant.

In related embodiments, the compositions of the invention furthercomprise an anti-fungal agent, an antibacterial agent, and/or collagen.In yet other related embodiments, the composition further comprises apharmaceutical carrier. In certain embodiments, the composition isformulated as a gel, solid, liquid, sponge, foam, spray, emulsion,suspension, or solution. In yet other related embodiments, thecomposition further comprises a neutral liquid, neutral gel or neutralsolid. In preferred embodiments, the composition further comprises aneutral solid and the neutral solid is a gauze. In other preferredembodiments, the composition is in the form of a coating on a neutralsolid. In yet other preferred embodiments the barrier-forming materialis a gauze.

The present invention provides for use of a coagulant with thecompositions, methods, and kits of the present invention describedherein, wherein the coagulant is selected from the group consisting ofalpha-2-antiplasmin, alpha-1-antitrypsin, alpha-2-macroglobulin,aminohexanoic acid, aprotinin, a source of Calcium ions, calciumalginate, calcium-sodium alginate, casein kinase II, chitin, chitosan,collagen, cyanoacrylates, epsilon-aminocaproic acid, factor XIII,fibrin, fibrin glue, fibrinogen, fibronectin, gelatin, living platelets,metha crylates, plasminogen activator inhibitor-1 (PAI-1), plasminogenactivator inhibitor-2 (PAI-2), plasmin activator inhibitor, plasminogen,platelet agonists, protamine sulfate, prothrombin, an RGD peptide,sphingosine, a sphingosine derivative, thrombin, thromboplastin, andtranexamic acid. In another embodiment of the invention, any coagulantthat can be used in any of the methods of the present inventiondescribed herein.

The present invention provides for use of a vasoconstrictor with thecompositions, methods, and kits of the present invention describedherein, wherein the vasoconstrictor is selected from the groupconsisting of endothelin, endothelin-1, epinephrine, adrenaline,metaraminol bitartrate (Aramine™), dopamine HCl (Intropine™),isoproterenol HCl (Isuprel™), norepinephrine (Levophed™), phenylephrine,Serotonin™, thromboxane, norepinephrine, prostaglandin, methergine,oxytocin, isopreland U-46619, papaverine, yohimbine, visnadin, khellin,bebellin, and nicotinate derivatives. In an alternative embodiment, thepresent invention provides for a vasoconstrictor that can be used withthe methods of the present invention described herein, with the provisothat the vasoconstrictor is not epinephrine.

In certain embodiments of the methods of the invention, the patient is ahuman.

In certain embodiments of the compositions, methods, and kits of theinvention, a film or membrane is used in conjunction with thevasoconstrictor and/or coagulant. In certain modes of thesesembodiments, the film or membrane comprises a barrier-forming material.In yet other embodiments, the composition is formulated as a mat,string, microbead, microsphere, and/or microfibril.

The methods and compositions of the present invention provide for acomposition that comprises a vasoconstrictor and or coagulant andfurther comprises a biodegradable material. In certain embodiments, thebiodegradable material is selected from the group consisting ofpolyanionic polysaccharides, alginic acid, collagen, polyglycolide,polylactide, polycaprolactone, dextran and copolymers thereof,polyglycolide, polylactide, polydioxanones, polyestercarbonates,polyhydroxyalkonates, polycaprolactone, and copolymers thereof. Incertain embodiments, the compositions of the invention are coated withone or more of these biodegradable materials. In other embodiments, thecompositions of the invention are co formulated with one or more ofthese biodegradable materials.

In preferred embodiments, the methods of the invention comprise the stepof administering to the patient an anticoagulant, and then applyingtopically to the patient's skin over a wound contiguous with the breachor puncture in the vein or artery a composition comprising an effectiveamount of a vasoconstrictor, wherein the vasoconstrictor does notcomprise a poly-β-1→4 N-acetylglucosamine polymer or derivative thereof,and concurrently carrying out the step of applying compression to thebreached or punctured vein or artery. In related preferred embodiments,the anticoagulant administered to a patient is selected from the groupconsisting of coumadin, heparin, nadroparin, aspirin, and a thrombolyticagent. In other related embodiments, the composition further comprisesprotamine sulfate in an amount effective to neutralize heparin. In otherpreferred embodiments, the patient is administered the anticoagulantcomprising aspirin together with another agent selected from the groupconsisting of coumadin, heparin, nadroparin, and a thrombolytic agent.In yet other preferred embodiments, the anticoagulant comprises one ormore of coumadin, heparin, nadroparin, aspirin, or a thrombolytic agent.

The methods described herein can be used to treat a breach or puncturein any vein or artery, including but not limited to the femoral, radial,brachial, or axillary artery, and the femoral, internal jugular, orsubclavian vein. Descriptions and depictions of the locations of suchblood vessels can be found in anatomy references including but notlimited to Leonard, C. H. 1983, The concise Gray's Anatomy, Chartwellbooks, Secaucus, N.J.

In certain preferred embodiments of the methods of the invention, thecompression that is applied is manual compression. In other embodimentsof the present invention, the compression is mechanical compression. Inrelated embodiments, the compression is applied to the vein or arteryproximal of the puncture or breach. In other related embodiments, thecompression is applied at the site of application of the composition. Inyet other embodiments, the compression is applied with a compressionbandage. In preferred embodiments of the present invention, the methodsof the invention further comprise, repeating the step of application ofcompression to the breached or punctured vein or artery.

In certain embodiments of the present invention, the rate of cessationor reduction of blood flow out of the breach or puncture in the vein orartery achieved by the present methods involving the topical use of avasoconstrictor is at least 10% greater than without thevasoconstrictor. In other embodiments of the present invention, the rateof cessation or reduction of blood flow out of the breach or puncture inthe vein or artery is 20%, 30%, 40%, or 50% greater than applyingcompression in conjunction with a topical barrier-forming materialwithout a vasoconstrictor. In preferred embodiments of the presentinvention, the vein or artery is breached or punctured by a catheter.

In certain embodiments of the present invention, the time to achievecessation or reduction of blood flow out of the breach or puncture inthe vein or artery is 10% less than applying compression in conjunctionwith a topical barrier-forming material without a vasoconstrictor. Inother embodiments of the present invention, the time to achievecessation or reduction of blood flow out of the breach or puncture inthe vein or artery is 20%, 30%, 40%, or 50% less than applyingcompression in conjunction with a topical barrier-forming materialwithout a vasoconstrictor. In preferred embodiments of the presentinvention, the vein or artery is breached or punctured by a catheter.

The methods of the invention can be used to treat vascular breaches orpunctures in a patient by applying a composition of the invention to aportion of the patient's skin that is contiguous with the breach orpuncture. The portion of the skin can be at any distance from the breachor puncture, as long as the composition of the invention is capable ofexerting an effect on the wound healing process. In certain embodiments,the distance is approximately 10, 9, 8, 7, 6, 5, 4, 3, 2 or 1 cm fromthe puncture in a vein or artery. In a preferred embodiment, the breachor puncture in a vein or artery is approximately 8, 9, or 10 cm from acontiguous skin wound site. In another preferred embodiment, the breachor puncture in a vein or artery is approximately 4, 3, 2, or 1 cm from acontiguous skin wound site. In yet another preferred embodiment, thebreach or puncture in a vein or artery is approximately 7, 6, or 5 cmfrom a contiguous skin wound site.

The present invention provides methods for decreasing the occurrence oflocalized vascular complications in a patient. One such method comprisesthe following steps, a) applying topically to the patient's skin over awound contiguous with a breach or puncture in a vein or artery acomposition comprising a vasoconstrictor or coagulant, wherein thevasoconstrictor or coagulant does not comprise a poly-β-1→4N-acetylglucosamine polymer or derivative thereof; b) concurrentlyapplying compression to the breached or punctured vein or artery, and c)recording the occurrence of localized vascular complications, wherein anamount of the vasoconstrictor or coagulant is effective to cause sealingof the breach or puncture in the vein or artery, reducing the rate oflocalized vascular complications in comparison to applying compressionin conjunction with a topical barrier-forming material without avasoconstrictor. In certain preferred embodiments, the rate of cessationor reduction of blood flow out of the breach or puncture in the vein orartery is at least 50% greater than applying compression in conjunctionwith a topical bather without a vasoconstrictor. In other preferredembodiments, the rate of cessation or reduction of blood flow out of thebreach or puncture in the vein or artery is at least 10%, 20%, 30%, 40%,50%, 60%, or 70% greater than applying compression in conjunction with atopical barrier without a vasoconstrictor. In certain preferredembodiments, the time to achieve cessation or reduction of blood flowout of the breach or puncture in the vein or artery is at least 50% lessthan applying compression in conjunction with a topical barrier withouta vasoconstrictor. In other preferred embodiments, the time to achievecessation or reduction of blood flow out of the breach or puncture inthe vein or artery is at least 10%, 20%, 30%, 40%, 50%, 60%, or 70% lessthan applying compression in conjunction with a topical barrier withouta vasoconstrictor.

The present invention further provides a pharmaceutical composition fortopically treating a breach or puncture in a vein or artery wherein thepharmaceutical composition comprises: a) an effective amount of avasoconstrictor, wherein the vasoconstrictor does not comprise apoly-β-1→4 N-acetylglucosamine polymer or derivative thereof; and b) apharmaceutically acceptable carrier. In certain preferred embodiments,the pharmaceutical composition further comprises a coagulant. In otherpreferred embodiments, the pharmaceutical composition further comprisesa neutral liquid, neutral gel and/or neutral solid. As used herein inrelation to all embodiments of the invention, a neutral liquid, neutralgel and/or neutral solid means such agents have no activity or areinactive with respect to processes involved in hemostasis. Thepharmaceutical composition can further comprise a gauze.

The present invention further provides a kit useful for practicing thepresent methods that comprises in one or more containers a compositionuseful for treating a breach or puncture in a vein or artery, thatcomprises: a) an effective amount of a vasoconstrictor, wherein thevasoconstrictor does not comprise a poly-β-1→4 N-acetylglucosaminepolymer or derivative thereof; and b) a pharmaceutically acceptablecarrier, and instructions for topically treating a breach or puncture ina vein or artery. The composition can further comprise a coagulant, aneutral liquid, a neutral gel and/or a neutral solid. In a preferredembodiment the composition further comprises a gauze. In anotherpreferred embodiment the composition further comprises gelatin.

The present invention yet further provides a pharmaceutical compositionfor topically treating a breach or puncture in a vein or artery whereinthe pharmaceutical composition comprises: a) an effective amount of avasoconstrictor, wherein the vasoconstrictor does not comprise apoly-β-1→4 N-acetylglucosamine polymer or derivative thereof; and b)wherein the vasoconstrictor is formulated into a barrier, membrane, orfilm. The pharmaceutical composition can further comprise a coagulant.The pharmaceutical composition can further comprise a neutral liquid,neutral gel and/or neutral solid. In a preferred embodiment thecomposition further comprises gelatin.

The present invention yet further provides a kit useful for practicingthe present methods that comprises in one or more containers acomposition that comprises: a) an effective amount of a vasoconstrictor,wherein the vasoconstrictor does not comprise a poly-β-1→4N-acetylglucosamine polymer or derivative thereof; and b) wherein thevasoconstrictor is formulated into a barrier, membrane, or film, andinstructions for topically treating a breach or puncture in a vein orartery. The composition can further comprise a coagulant, a neutralliquid, a neutral gel and/or a neutral solid. In a preferred embodimentthe composition further comprises gauze. In another preferred embodimentthe composition further comprises gelatin.

In preferred embodiments, the compositions and methods of the inventiondescribed herein can be used in treatment of catheter-induced puncturesin veins or arteries. The treatment can comprise reducing hemostasistime, reducing the flow of blood from the catheter puncture in the bloodvessel, and reducing the occurrence of localized vascular complications.

In particular, the methods of the present invention described herein areuseful for the treatment of cardiac catheterization wounds. Such woundscommonly result from diagnostic or therapeutic cardiac interventionprocedures such as coronary angiography and angioplasty. In oneembodiment, a method for treating a puncture in a vein or arteryresulting from a cardiac catheterization procedure in a patientcomprises: a) applying topically to the patient's skin over a catheterexit site a composition comprising an effective amount of avasoconstrictor and/or coagulant, wherein the vasoconstrictor and/orcoagulant does not comprise a poly-β-1→4 N-acetylglucosamine polymer orderivative thereof, and wherein the catheter exit site is contiguouswith the catheter puncture in the vein or artery by approximately 1-10cm; and concurrently b) applying compression to the punctured vein orartery, wherein a cessation or reduction of blood flow out of the breachor puncture in the vein or artery is achieved at a rate 30%-50% greaterthan applying compression in conjunction with a topical barrier-formingmaterial without a vasoconstrictor and/or coagulant.

In another embodiment, a method for treating a puncture in a vein orartery resulting from a cardiac catheterization procedure in a patientcomprises: a) applying topically to the patient's skin over a catheterexit site a composition comprising an effective amount of avasoconstrictor and/or coagulant, wherein the vasoconstrictor and/orcoagulant does not comprise a poly-β-1→4 N-acetylglucosamine polymer orderivative thereof, and wherein the catheter exit site is contiguouswith the catheter puncture in the vein or artery by approximately 1-10cm; and concurrently b) applying compression to the punctured vein orartery, wherein a cessation or reduction of blood flow out of the breachor puncture in the vein or artery is achieved in a measure of time thatis 30%-50% less than applying compression in conjunction with a topicalbarrier-forming material without a vasoconstrictor and/or coagulant.

The methods of the present invention are also useful for reducing thenumber and size of hematomas such as those resulting from a cardiaccatheterization procedure in a patient. The methods of the presentinvention are also useful for inhibiting the formation of hematomas suchas those resulting from a cardiac catheterization procedure in apatient. As used herein in the context of the invention, “inhibiting theformation of hematomas” means decreasing the probability that a hematomawill form or decreasing the probability that hematomas of a large size,e.g., greater than 3 cm, will form. In one embodiment, a method ofreducing the occurrence of a hematoma following cardiac catheterizationcomprises: a) applying topically to the patient's skin over a catheterexit site contiguous by 1-10 cm with a catheter puncture in a vein orartery a composition comprising an effective amount of a vasoconstrictorand/or coagulant, wherein the vasoconstrictor and/or coagulant does notcomprise a poly-β-1→4 N-acetylglucosamine polymer or derivative thereof;b) concurrently applying compression to the punctured vein or arterythereby reducing the occurrence of hematomas after cardiaccatheterization. In certain embodiments, a reduction in the occurrenceof hematomas is measured by recording the number of hematomas formed ina sample patient population, in comparison to a sample patientpopulation wherein compression is applied to the patients in conjunctionwith a topical barrier-forming material without a vasoconstrictor and/orcoagulant.

3.1 Definitions

“Localized vascular complications”—Localized vascular complicationsinclude hematomas, pseudoaneurysms, and AV fistula formations, and othermedical conditions associated with a breach or puncture in a vein orartery.

“Hematoma”—Hematomas are produced by of the invasion of other tissues byblood tissue normally localized inside the blood vessels, resulting inswelling of the invaded tissues and tissues surrounding the invadedtissues.

“Barrier-forming material”—A barrier-forming material is any materialthat can be placed on the surface of the skin over a wound andphysically impede or reduce blood flow from the wound.

“Concurrently”—concurrently as used herein, in relation to applying acomposition of the invention and compression to a patient, means eitherapplying compression and a composition of the invention topically to awound site on a patient at the same time, or applying a compositiontopically to a wound site on a patient immediately (i.e. any period oftime less than 1 minute, or less than 2 minutes) followed by applicationof compression to the wound site on a patient. For example, if thecomposition of the invention is formulated as a barrier formingmaterial, then the composition and the compression can be applied at thesame time. If the composition is for example a gel, the gel can beapplied to the wound site followed immediately by compression, or thegel might be applied to a barrier-forming material such as gauze and thetreated gauze can then be applied at the same time as compression. Inall examples the application of the composition and compression isdefined as concurrent.

4. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1. Ultrasonography of a human femoral artery. Top of imagecorrelates with skin surface of patient, and cross hair symbol marks thecenter of the blood vessel at 2.67 cm below the skin surface.

5. DETAILED DESCRIPTION OF THE INVENTION

The present invention is based upon the concept of treating a breach orpuncture in a vein or artery from a distance via a skin surface woundcontiguous with the breach or puncture. The present methods encompasstopically applying to the skin surface wound a composition withvasoconstrictive and/or coagulating properties in conjunction withapplication of compression to the site or an proximal site of the veinor artery to reduce blood flow. The methods described herein seal orheal of the breach or puncture at a greater rate or in less time incomparison to available non-invasive treatments, i.e. hemostaticpatches, compression. The invention also encompasses one or morecompositions that can be used in the methods of the invention and kitsthat comprise one or more of the compositions of the invention for usein treating a breach or puncture in a vein or artery.

Thus, the present invention provides a solution for the problem oflocalized vascular complications, such as hematomas, pseudoaneurysms,and AV fistula formations, associated with treating a breach or puncturein a vein or artery. In particular, the methods of the inventiondescribed herein, are an improvement over both hemostatic patches andarterial closure devices in treating wounds resulting from medicalprocedures, including but not limited to cardiac catheterizationprocedures. Whereas the available hemostatic patches act directly at thesite of contact with the wound, and only indirectly act on the catheterpuncture in the artery or vein by decreasing blood flow, without beingbound by any theory, the inventors believe that, the compositions andmethods of the invention act on the catheter puncture in the artery orvein from a distance and stop or reduce blood flow from the internalpuncture site at a greater rate or in less time than presently knowndevices, decreasing the likelihood of hematoma formation. For example asoluble vasoconstrictor may act on receptors present in a blood vessel,resulting in a constriction of the blood vessel. The methods of thepresent invention are an improvement over the invasive arterial closuredevices used following cardiac catheterization. The advantages offeredinclude the ease of carrying out the method steps, since no suturemachines or application devices need be applied subcutaneously, and adecreased likelihood of hematoma formation. The non-invasive methods ofthe invention also minimize the risk of infections that can occur whenforeign objects, such as the available arterial closure devices areinserted through the layers of a patients skin.

5.1 Injuries for Treatment by the Present Methods and Compositions

The present invention provides methods for treatment of a breach orpuncture in a vein or artery. The breach or puncture may be a wound ofany origin or cause. In an embodiment of the invention, the breach orpuncture in a vein or artery is caused by an object such as a bullet,knife, or surgical instrument. In a preferred embodiment the object thatcauses a breach or puncture in the vein or artery is a catheter. In yetanother preferred embodiment the puncture is the result of a cardiaccatheterization procedure. Arteries whose breaches or punctures may betreated by the present methods and compositions include, but are notlimited to, the femoral, radial, brachial, and axillary arteries. Veinswhose breaches or punctures may be treated by the present methods andcompositions include, but are not limited to, the femoral, internaljugular, and subclavian veins.

Generally, when the breach or puncture in the vein or artery beingtreated by the present methods and compositions results from cardiaccatheterization, the vein or artery will have a breach or puncture canbe caused by a smaller diameter catheter. Examples of smaller diametercatheters include catheters ranging from 4 F to 6 F in size (F=FrenchUnits, where 1 F is equal to 0.33 mm). In another embodiment of theinvention, the breach or puncture may be caused by a larger-diametercatheter ranging in size from 7 F to 11.5 F (Sanborn et al., 1993,Journal of the American College of Cardiology. 22(5):1273-1279).Preferably, the artery is larger than a catheter between 6 F and 8 F insize. In such embodiments where a breach or puncture is the result ofcatheterization, the methods and compositions of the invention can beused to treat punctures or breaches cause by various types of catheters,including but not limited to a Swan-Ganz catheter, a Sones catheter, anda pigtail catheter (Olade et al., 2002, Emedicine Journal, 3(3):1-13).

In certain embodiments, catheters are inserted perpendicular to the skinsurface. In other embodiments, a catheter is inserted at an angle thatis 20°-40° to the skin surface.

The present methods can be used to treat breaches or punctures in veinsor arteries in any mammal including but not limited to a human, dog,horse, cat, rabbit, rat, mouse, pig, cow, monkey or sheep. Preferablythe patient is a human.

5.2 Methods for Using the Hemostatic Compositions

The methods of the invention encompass topical administration of acomposition comprising a vasoconstrictor or a coagulant to a patient'sskin surface at a site of a wound that is contiguous with a breach orpuncture in a vein or artery. Compositions of the invention comprisingvasoconstrictors and/or coagulants are further described in section 5.5below, and may include a barrier-forming component that is applied tothe skin surface over a wound that is contiguous with a breach orpuncture in a vein or artery.

5.2.1 Site of Application of Hemostatic Composition

The present methods for treating vascular wounds can entail acombination of applying pressure and/or a barrier-forming material and acomposition of the invention comprising a vasoconstrictor or coagulantagent to a site on the patient's skin surface wound that is contiguouswith a breach or puncture in a vascular structure such as a vein orartery. In a preferred embodiment of the invention, a site ofapplication on the skin surface of the composition of the invention,contiguous with a breach or puncture in the vein or artery, is within 10cm of the site of a breach or puncture in a vein or artery. In anotherembodiment, the site of application is within approximately 9 cm, 8 cm,7 cm, 6 cm, 5 cm, 4 cm, 3 cm, 2 cm, or 1 cm of a breach or puncture inthe vein or artery. In yet another embodiment, the site of topicalapplication is between 10 cm and 2 cm of a breach or puncture in thevein or artery. In a preferred embodiment the site of topicalapplication is about 1-10 cm from a breach or puncture in the vein orartery.

5.2.2 Applying Compression

Compression can be indirectly applied to a breached or punctured vein orartery in several ways. The present methods for treating vascular woundscan entail a combination of applying pressure and/or a barrier-formingmaterial and a composition of the invention comprising a vasoconstrictoror coagulant agent to a site on the patient's skin surface wound that iscontiguous with a breach or puncture in a vascular structure such as avein or artery. When a vein or artery with a breach or puncture issubject to compression, blood flow and pressure decreases in the vein orartery, allowing the puncture to seal or clot with a decreasedlikelihood of hematoma formation.

In certain embodiments of the invention, the compression is manualcompression. Manual compression can be applied by pressing with the tipsof fingers on the skin surface, at a point above an underlying vein orartery, so that the vein or artery is compressed and blood flow issignificantly reduced or stopped in the targeted vein or artery. Thistype of compression can be applied at a site proximal of a breach orpuncture in a vein or artery. Typically, an proximal site is between 1and 10 cm proximal of the breach or puncture. The technique of manuallypressing on a patients skin at the correct place to reduce blood flowand blood pressure is common in the art and is effective in both humansand other mammals (Camenzind et al, 1994, Journal of the AmericanCollege of Cardiology. 24(3):655-662; Kipshidze et al., 1998, Journal ofInvasive Cardiology. 10(3):133-141; Merino et al., 1992, Catheterizationand Cardiovasular Diagnosis 26:319-322; Sanborn et al., 1993, Journal ofthe American College of Cardiology. 22(5):1273-1279). The manualcompression can be optionally applied in combination with application ofa composition comprising “a composition of the invention,” comprising avasoconstrictor or coagulant. Such embodiments entail manuallycompressing to subject a artery or vein to compression concurrently withapplying a composition of the invention. Alternatively, manualcompression can be applied immediately prior to application of acomposition of the invention or immediately after application of acomposition of the invention.

In an embodiment of the invention that encompasses a manual compressiontechnique as a part of a method of the invention, the technique may beone wherein pressure is applied with ones fingers or the palm of oneshand directly over a dermotomy site that is contiguous with a breach orpuncture in the vein or artery (Sanborn et al., 1993, Journal of theAmerican College of Cardiology. 22(5):1273-1279). In certain embodimentswhere the wound is caused by a catheter, compression can be applieddirectly to the wound site at the same angle that the catheter wasinserted at. In one embodiment of the invention, manual compression maybe applied concurrently with application of the composition. If thecomposition of the invention is formulated as a harrier-formingmaterial, then the composition and the compression can be applied at thesame time. If the composition is for example a gel, the gel can beapplied to the wound site followed immediately by compression, or thegel might be applied to a barrier-forming material such as gauze and thetreated gauze can then be applied at the same time as compression.

In applying manual compression techniques, the amount of pressureapplied and the consistency of pressure force generally varies over timeand among individuals applying the pressure. For this reason, it may bepreferable to apply compression to a breached or punctured vein orartery mechanically rather than manually, to ensure the consistency ofthe force of the pressure applied. Several devices have been developedand are commonly used to apply compression, including C-clamps ofvarying or adjustable force and compression bandages/dressings withattachment of desired weights. Such compression devices may be used inconjunction with the methods and compositions of the invention and canprovide a constant application of the desired compression force.

In certain embodiments, compression is either applied first at a siteproximal to the wound site to decrease blood flow in the vein or arterywith the breach or puncture or compression is first applied to both theproximal site and the wound site simultaneously, in either case thecompression applied at the proximal site is later released or decreasedwhile the compression at the wound site is maintained. The time betweenthe application of compression and the release of pressure at theproximal site can be about 15 seconds, 30 seconds, 1 minute, 1.5minutes, 2 minutes, 2.5 minutes, 3 minutes, 3.5 minutes, 4 minutes, 4.5minutes, or 5 minutes. In other embodiments, compression is applied andmaintained at both the proximal and wound sites.

In another embodiment, where the composition encompasses an adhesivebarrier-forming material, the composition can be topically adhered tocover a skin surface wound that is contiguous with a breach or puncturein a vein or artery without applying pressure to compress the vein orartery.

5.2.3 Compression Time

One of the advantages of the methods of the present invention is thatthe combination of compression and the compositions of the inventionresults in a reduction in compression time needed to stop or slow theflow of blood from the breach or puncture relative to compression alone.The amount of time for which compression is maintained to achievehemostasis, i.e. a cessation or lessening of blood flow from a breach orpuncture in a vein or artery, is subjected to the size of the wound, thedistance from the skin surface, and the amount of pressure applied incompressing the blood vessel. As used herein in connection with theinvention, hemostasis means cessation of blood flow from a breach orpuncture in a vein or artery. The amount of time for which compressionis maintained when carrying out the methods of the invention is shorterin comparison to compression alone or compression without a compositionof the invention as measured under comparable circumstances.

In general, when applying a manual compression technique it ispreferable to apply adequate but gradually lessening pressure for aperiod of time between ten and thirty minutes before inspection of thesite, with reapplication of pressure as necessary for ten to twentyminute intervals (Camenzind et al, 1994, Journal of the American Collegeof Cardiology. 24(3):655-662; Kipshidze et al., 1998, Journal ofInvasive Cardiology. 10(3):133-141; Sanborn et al., 1993, Journal of theAmerican College of Cardiology. 22(5):1273-1279). In certain instances,the period of time for which compression is applied may be greater than30 minutes. If non-manual compression is employed in the methods of theinvention, then a constant pressure may be applied for ten to thirtyminutes or more, particularly when applying compression alone. Atechnique corresponding to stepwise reduction in compression describedabove can be employed when using for example a C-clamp or compressionbandage/dressing, wherein the pressure may be lessened in incrementsover time as necessary. A fluid-filled balloon connected to a pressuretransducer can also be used to measure the amount of pressure applied.The amount of compression force varies for each patient and the type andlocation of the blood vessel, i.e. vein or artery, being treated. Any ofthe compression techniques described herein can be used in carrying outthe methods of the invention, as well as other standard compressiontechniques.

In contrast, the time for applying compression, manual or otherwise, inconjunction with a composition of the invention is shorter and may bedivided into intervals at which to release or partially release pressurefor 1-10 seconds in order to record observations to calculate thecessation rate or time of blood from a puncture or breach and/or tocalculate the time to seal a puncture or breach. The time intervals maybe 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, or 60 seconds in length.Alternatively, the time intervals may be 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5,5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10 minutes in length (Silberet al., 1998, Catheterization and Cardiovasular Diagnosis 43:421-427).

In a related embodiment, compression is applied for time intervals ofequivalent length in patients receiving the treatment compression aloneand those receiving the treatment of compression in conjunction with thecomposition of the invention.

In an embodiment, the decrease in the amount of time required to achievecessation or a decrease in blood flow from a breach or puncture in avein or artery using the methods of the invention in comparison to usingcompression alone is quantified as a percentage.

Various methods for measuring compression time and compression force canbe employed in carrying out the methods of the invention as described insection 5.2.4.

Compression should be applied to the desired vein or artery shortlyfollowing the breach or puncture (or immediately following a breach orpuncture caused by a medical procedure), or as soon possible in the caseof accidental punctures or breaches. For example in one embodiment,caused by a medical procedure, compression is applied immediately afterremoval of a catheter and a catheter sheath.

In a preferred embodiment of the invention, one or more of the methodsencompassed by the invention described herein are effective at achievinga cessation or reduction of blood flow out of a breach or puncture in anartery or vein in 3, 4, 5, 6, 7, 8, 9, or minutes.

The compositions of the invention described herein in section 5.5 arepreferably applied concurrently with an initial application ofcompression. In other embodiments, the compositions can be reapplied atcompression time intervals. In other embodiments, compression is appliedfor a brief time prior to application of a composition of the inventionand further compression.

5.2.4 Measurement of Time or Rate of Cessation of Bleeding or Sealing ofBreach or Puncture

In one embodiment, the rate or time of cessation or reduction of bloodflow from a breached or punctured blood vessel achieved using thepresent methods can be measured by any technique known to those skilledin the art. In certain embodiments, for example when testing theefficacy of the present compositions, the rate or time of cessation orreduction is measured and compared in the absence or presence of acomposition of the invention. Preferably, the same technique is utilizedwhen measuring the rate or time of cessation or reduction in thepresence or absence of a composition of the invention.

In certain embodiments, the efficacy of the compositions and methods ofthe invention can be measured by simulating in dogs the cardiaccatheterization procedure used in humans. The femoral artery in dogs isparticularly desirable for such simulations since it is easilyaccessible, being located not far beneath the skin surface, andhematomas can be easily visualized in this region. Examples of cardiaccatheterization simulations in dogs and determination in such dogs ofrate or time of cessation of bleeding are presented in Example 1 ofSection 6. Measuring rate or time till cessation of blood flow from askin wound contiguous with a breach or puncture in a vein or artery isan indirect measurement of the flow of blood from the breach or puncturein the vein or artery. Additional, more direct, techniques that can beused with the methods of the invention described herein to observecessation of blood flow from the breach or puncture in the vein orartery are taught below in Section 5.3.

The rate of or time to achieve cessation or reduction of blood flow outof the breach or puncture in a vein or artery can be calculated simplyas the total number of time intervals required to achieve the effect,provided the time intervals are equivalent for each treatment, i.e.compression alone or compression in conjunction with the composition ofthe invention. For example if the sum of the number of time intervalsrequired for all patients receiving a first treatment is 8 and if thesum of number of time intervals required for all patients receiving asecond treatment is 10, then the percent difference would be 25%.Alternatively, the average time to achieve cessation could be calculatedfor each treatment group, then compared to determine the percentdifference.

In another embodiment, the time to achieve cessation is measured inminutes and/or seconds rather than time intervals. The percentdifference between the times measured can be calculated with one of themethods described above for the time intervals method.

Where the time to achieve cessation of blood flow from the breach orpuncture in the vein or artery is are measured in minutes and/orseconds, the number of time intervals for which compression is appliedmay not necessarily be a factor in calculating the time to cessation.Total time from application of treatment to cessation or reduction ofblood flow out of breached or punctured veins or arteries can becompared for the two treatment groups. In this embodiment of theinvention, compression alone may be initially applied for a longerperiod of time than compression in conjunction with the composition ofthe invention, since values for the time to achieve cessation are knownin the art for compression alone. The time to achieve cessation or therate of cessation of blood flow from the breach or puncture in the veinor artery can also be measured for the circumstance where onlycompression is applied. These measurements can be made by any of themethods described herein, including but not limited to ultrasonographyand scintigraphic imaging. For example, compression alone may be appliedto a patient for 10 minutes before an observation is recorded, while thepatient receiving compression in conjunction with the composition of theinvention would require compression to be partially and briefly releasedto make blood flow observations, in order to obtain data to calculatethe time to achieve cessation.

If a set number of compression timer intervals is applied, the percentdifference can be calculated based on the number of patients in eachtreatment group. For example, if compression is applied for two timeintervals and 60 of 100 patients receiving a first treatment consistingof compression alone achieve cessation of bleeding from a breach orpuncture or sealing of a breach or puncture in a vein or artery aftertwo compression intervals, and 80 of 100 patients receiving a secondtreatment consisting of compression plus the composition of theinvention achieve cessation or sealing of a breach or puncture after twoequivalent time intervals, then the percent difference in time toachieve hemostasis would be 20%.

The rate of hematoma occurrence can be calculated with clinicalobservations. One indicator of hematoma formation is swelling of theskin around the local area of a punctured or breached vein or artery.Alternatively, diagnoses of hematoma or local vascular complications aswell as sealing and rates or time to achieve cessation of blood flowfrom a breach or puncture may also be made with various imagingtechniques.

Statistical methodology can be employed to determine if the observeddifferences are statistically significant. In particular, thedifferences observed between the rate of cessation or time to achievecessation or occurrence of vascular complications using the methods ofthe invention in comparison to compression alone or compression withvasoconstrictors and/or coagulants that do not function at a distancecan be analyzed with standard statistical methodology. Statisticalsignificance can be determined with any standard calculated statistic(e.g., a one-tailed t-statistic, a two-tailed t-statistic, a chi-squarestatistic, an F-statistic, etc.). Standard statistical methodologiessuitable for use in connection with the invention include methodologiescommonly used in medical analyses and clinical trials. Examples ofmethodologies can be found in reference publications including but notlimited to: Vogt W., 1998, Dictionary of Statistics and Methodology-2nded., SAGE Publications; Spiegel, D., J. Myles, and K. R., 2002, AbramsBayesian Approaches to Clinical Trials and Health Care: Statistics inPractice. Wiley, John & Sons, Incorporated; Cleophas, T. J., A. H.Zwinderman, and T. F. Cleophas, 2002, Statistics Applied to ClinicalTrials-2nd ed., Gehan, E. A. A. and N. A. Lemak, 1994, Kluwer AcademicPublishers; and Statistics in Medical Research: Developments in ClinicalTrials-1st ed., Kluwer Academic Publishers.

In a preferred embodiment of the invention, the time to achievecessation or reduction of blood flow or the rate of cessation orreduction of blood flow out of a breach or puncture in the vein orartery is achieved at a greater rate or in less time than applyingcompression in conjunction with a topical barrier-forming materialwithout a vasoconstrictor or coagulant, and without the formation of ahematoma. In another embodiment, the time or rate of cessation orreduction of blood flow out of a breach or puncture in the vein orartery is achieved at a greater rate or in less time than applyingcompression in conjunction with a topical barrier-forming materialwithout a vasoconstrictor or coagulant, and the rate of hematomaoccurrence decreases.

For example, color flow duplex sonography of a puncture site can be usedto test for pseudoaneurysms, and AV fistulas (Gwechenberger et al, 1997,Angiology. 48(2):121-126.) Scintigraphic Image Analysis can be employedto examine bleeding and/or hematoma formation at the site of breach orpuncture and to determine if a breach or puncture is effectively sealed(Ismail et al., 1995, Catheterization and Cardiovasular Diagnosis34(1):88-95). Angiogram technology can also be used to examine sealingof a breach or puncture site (Hoekstra et al., 1998, Biomaterials.19:1467-1471). With imaging analyses as described herein and in theexamples section, the time or rate and percent difference in rates ortimes of cessation or reduction of blood flow out of a breach orpuncture in the vein or artery can be calculated without releasingcompression to make observations.

In embodiments where the femoral artery is punctured, distal pulsemeasurements can be used to determine if effective closure of thepuncture has been achieved. Similar techniques can be used on otherarteries or veins.

In one aspect of the invention, the rate of cessation or reduction ofblood flow out of a breach or puncture in the vein or artery whenemploying the methods and compositions of the invention is 10% greaterthan applying compression in conjunction with a topical barrier or gauzewithout a vasoconstrictor or coagulant. In a related aspect of theinvention, the rate of cessation or reduction of blood flow out of abreach or puncture in the vein or artery when employing the methods andcompositions of the invention is 10% greater than applying compressionin conjunction with a topical barrier or gauze comprising less than aneffective amount of a vasoconstrictor. In other aspects, the rates asdescribed above are 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%,70%, 80% or greater.

In one aspect of the invention, the time to achieve cessation orreduction of blood flow out of a breach or puncture in the vein orartery when employing the methods and compositions of the invention is10% less than applying compression in conjunction with a topical barrieror gauze without a vasoconstrictor or coagulant. In a related aspect ofthe invention, the time to achieve cessation or reduction of blood flowout of a breach or puncture in the vein or artery when employing themethods and compositions of the invention is 10% less than applyingcompression in conjunction with a topical bather or gauze comprisingless than an effective amount of a vasoconstrictor. In other aspects,the time as described above is 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%,50%, 60%, 70%, 80% or less.

In a preferred embodiment, the methods and compositions of the inventioneffectively cause a cessation of blood flow from both a breach orpuncture site in a vein or artery and from the wound site at the skinsurface.

In a another preferred embodiment, the methods and compositions of theinvention effectively cause a cessation of blood flow from both a breachor puncture site, cause a cessation of blood flow from the wound site atthe skin surface, and decrease the likelihood of localized vascularcomplications including hematoma formation.

5.2.5 Scintigraphic Image Analysis

Scintigraphic Image Analysis can be employed to examine bleeding and/orhematoma formation at site of breach or puncture and to determine if abreach or puncture is effectively sealed, especially in animal modelsystems (Ismail et al., 1995, Catheterization and CardiovasularDiagnosis 34(1):88-95). For example if the invention is being tested onthe femoral arteries of dogs, a camera fitted with a low energyall-purpose collimator can be placed in position over a puncture orbreach site. Point sources (˜50 μCi 99 mTc) in suitable containers maybe used to record the exact position of the leg, for alignment ofsubsequent images. Dogs are injected through the external jugular venouscannula with autologous technetium-labeled red blood cells,(mean=16.65±3.51 mCi), for which the labeling efficiency is previouslytested. The labeled cells are allowed to circulate for 5 minutes beforeimages are taken. Images can be taken at regular time intervals up to 24hours. Early images can be used to ascertain sealing of a breach orpuncture and cessation of blood flow from the breach or puncture,whereas later images can be used to identify hematomas with greatefficiency.

5.2.6 Angiography

An angiogram is a test in which a dye, or contrast, is administered to apatient and X-ray imaging is used to look inside blood vessels in orderto diagnose or treat lesions involving the blood vessels. Angiogramtechnology is commonly employed to guide catheters through the femoralartery in cardiac catheterization procedures where the femoral artery ispunctured. Angiograms can also be used to examine, in a non-invasivemanner, cessation of blood flow from a breach or puncture in a vein orartery, or sealing of such a breach or puncture (Hoekstra et al., 1998,Biomaterials. 19:1467-1471). CT or MRI imaging can also be used toexamine sealing of a breach or puncture site and cessation of blood froma breach or puncture in a vein or artery.

5.2.7 Ultrasonography

Various sonography and ultrasonography techniques may be employed withthe methods and compositions of the invention to examine bleeding and/orhematoma formation at site of breach or puncture and to determine if abreach or puncture is effectively sealed. FIG. 1 exemplifies another useof ultrasonography, determining the distance between the skin surfaceand an underlying vein or artery in a human. Duplex ultrasonography hasdemonstrated useful clinical applications in peripheral arterial testingfor lesion localization and quantification of abnormal blood flow. Forexample, color flow duplex sonography of a puncture site can be used totest for hematoma, pseudoaneurysm, and AV fistula formation(Gwechenberger et al, 1997, Angiology. 48(2):121-126).

5.2.8 Thromboelastography

Thromboelastography is a means of examining the whole process of bloodcoagulation using whole blood. Thromboelastography is useful inconnection with the present invention to determine time to cessation orreduction of blood flow from a breached or punctured vein or artery. InThromboelastography measurements of time and clot strength are made. Theresults of the measurements are typically presented in a graphrepresenting the beginning of clot formation to fibrinolysis. The graphrepresents the time necessary for a clot to form and the tensilestrength of a clot. A clot has both viscous and elastic properties,however a thromboelastograph measures only the elastic properties of theclot. Typically clotting time is plotted on the x-axis and clot firmnessor elasticity is plotted on the y-axis. The clotting time can beeffected by various factors including clotting factor deficiencies,inhibitors, anticoagulants, and low platelets. Graphed measurementstypically result in an ovoid shape. The alpha angle is measured betweenthe vertical midline of the ovoid shape to the beginning of clottingtime. The alpha angle and the clot formation time indicate how fast theclot structure is forming, which can be effected by clotting factordeficiencies, platelet dysfunction, thrombocytopenia,hypofibrinogenaemia. Maximum clot firmness is defined as the width ofthe curve at the widest point. Maximum clot firmness can be effected bylow platelets or low figbrinogen. Fibrinolysis is measured as a decreasein amplitude from the maximum. Typically a decrease of 15% or more is anindication that fibrinolysis is taking place. Examples ofthromboelastographs that can be used in conjunction with the claimedinvention include but are not limited to ROTEG™, and TEG(Tromboelastography Coagulation Analizer, Haemascope corporation, Niles,Ill.). A vasoconstrictor and/or coagulant of the invention can beexamined by measuring the time to form a clot and the strength of a clotsealing a breach or puncture in a vein or artery withthromboelastography. The speed and strength of clotting can be used indetermining if a breach or puncture in a vein or artery is effectivelysealed.

5.2.9 Administration of Anticoagulants

In certain embodiments of the invention, the methods and compositions ofthe invention may be administered to a patient to whom an anticoagulantin an effective amount to prevent coagulation of blood has beenadministered. Examples of anitcoagulants that may be used in conjunctionwith the invention include, Coumadin™, Dicumarol™, heparin, nadroparin,aspirin, an antiplatelet drug, or a thrombolytic agent. Typically, fulldose of heparinization for more than 12 hours before a medical procedureis common. In other protocols heparin is administered at repeatedintervals to ensure that a constant active clotting time is maintained(Falstrom et al., 1997, Catheterization and Cardiovasular Diagnosis41:79-84). For those patients to whom an anticoagulant has beenadministered prior to a puncture or breach in a vein or artery, thecomposition of the invention may further comprise one or more agentsthat locally neutralize the effect of the administered anticoagulant. Inpatients to whom heparin has been administered the composition mayfurther comprise protamine sulfate in an amount effective to locallyneutralize heparin.

5.3 Pharmaceutical Compositions

The present invention provides pharmaceutical compositions that can beused in the practice of the methods of the invention. The pharmaceuticalcompositions encompassed by the present invention comprise an effectiveamount of one or more vasoconstrictors and/or one or more coagulants.The vasoconstrictors and/or coagulants that can be used in the methodsof the invention can be any vasoconstrictor and/or coagulant thatfunctions at a distance. The compositions of the invention can beformulated into a wide variety of shapes, sizes and masses. Thecompositions can additionally include components that have no knownvasoconstrictive or coagulant properties.

As used herein, the term “effective amount” means a quantity of acomposition comprising a vasocontrictor and/or coagulant that can beapplied topically and function, as a vasoconstrictor and/or a coagulant,at a distance to treat a breach or puncture in an artery or vein. Theeffectiveness of the amount can be measured by comparing the time toachieve or rate at which blood flow from a breach or puncture in a veinor artery ceases or slows in comparison to compression alone.

In certain embodiments of the invention, the compositions of theinvention can comprise combinations of agents in the methods of theinvention. In one embodiment, the combination can comprise avasoconstrictor and a coagulant. In other embodiments, the compositioncan comprise a greater quantity of vasoconstrictor than coagulant, agreater quantity of coagulant than vasoconstrictor. In yet otherembodiments, the compositions comprise an agent that functions as both avasoconstrictor and coagulant. The term “effective amount,” as usedherein, also means a total amount of a composition of the invention thatis applied in the methods of the invention.

The size, shape, and mass of the composition used to topically treat abreach or puncture in a vein or artery will vary depending on theparticular use. A pharmaceutical composition can contain an effectiveamount of a vasoconstrictor, which unless otherwise indicated, does notcomprise a poly-β-1→4 N-acetylglucosamine polymer or derivative thereof.A pharmaceutical composition of the invention can also contain apharmaceutically acceptable carrier.

As used herein derivatives of a poly-β-1→4 N-acetylglucosamine polymerinclude: a semi-crystalline form of a poly-β-1→4-acetylglucosaminepolymer; a poly-β-1→4-acetylglucosamine polymer comprising about 50 toabout 150,000 N-acetylglucosamine monosaccharides covalently attached ina β-1→4 conformation, and said polymer has a molecular weight of about10,000 daltons to about 30 million daltons; apoly-β-1→4-acetylglucosamine polymer comprising about 50 to about 50,000N-acetylglucosamine monosaccharides covalently attached in a β-1→4conformation, and said polymer has a molecular weight of about 10,000daltons to about 10 million daltons; a poly-β-1→4-acetylglucosaminepolymer comprises about 50 to about 10,000 N-acetylglucosaminemonosaccharides covalently attached in a β-1→4 conformation, and saidpolymer has a molecular weight of about 10,000 daltons to about 2million daltons; a poly-β-1→4-acetylglucosamine polymer comprising about50 to about 4,000 N-acetylglucosamine monosaccharides covalentlyattached in a β-1→4 conformation, and said polymer has a molecularweight of about 10,000 daltons to about 800,000 daltons; and asemi-crystalline poly-β-1→4 N-acetylglucosamine polymer comprising atleast one N-acetylglucosamine monosaccharide that is deacetylated, andwherein at least 40% of said N-acetylglucosamine monosaccharides areacetylated.

Derivatives of a poly-β-1→4 N-acetylglucosamine polymer also includecompositions that are 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%or less poly-β-1→4 N-acetylglucosamine. Thus, with varying degrees ofpurity of a vasoconstrictor and/or coagulant is encompassed by theinvention.

In certain embodiments of the invention, the composition of theinvention comprises a vasoconstrictor and/or a coagulant with one ofmore of the following provisos that the composition of the inventiondoes not comprise in certain embodiments:

1. The composition of the invention does not comprise a poly-β-1→4N-acetylglucosamine polymer or derivative thereof. The composition doesnot comprise a poly-β-1→4 N-acetylglucosamine polymer or derivativethereof and an agent that functions as a vasoconstrictor and/orcoagulant

2. The composition of the invention does not comprise a poly-α-1→4N-acetylglucosamine polymer or derivative thereof. The composition doesnot comprise a poly-α-1→4 N-acetylglucosamine polymer or derivativethereof and an agent that functions as a vasoconstrictor and/orcoagulant

3. The composition of the invention does not comprise a poly-α-1→4N-acetylglucosamine polymer or derivative thereof and a poly-β-1→4N-acetylglucosamine polymer or derivative thereof. The composition doesnot comprise a poly-α-1→4 N-acetylglucosamine polymer or derivativethereof, poly-β-1→4 N-acetylglucosamine polymer or derivative thereof,and an agent that functions as a vasoconstrictor and/or coagulant

4. The composition of the invention does not comprise chitosan, achitosan derivative, or compositions natural or synthetic that comprise10%400% chitosan or a chitosan derivative. The composition of theinvention does not comprise chitosan or a chitosan derivative derivedfrom crustaceans. The composition of the invention does not comprisechitosan or a chitosan derivative derived from crustaceans wherein thechitosan or a chitosan derivative have been purified or modified.

5. The composition does not comprise chitosan or a chitosan derivativethat is 80% or more deacetylated. The composition does not comprisechitosan or a chitosan derivative that is 70%-100% deacetylated.

6. The composition does not comprise chitosan or a chitosan derivativethat is free of fungal, micro-fungal, and crustacean chitosan orchitosan derivatives.

7. The composition of the invention does not comprise chitin, a chitinderivative, or compositions natural or synthetic that comprise 10%-100%chitin or a chitin derivative. The composition of the invention does notcomprise chitin or a chitin derivative derived from crustaceans. Thecomposition of the invention does not comprise chitin or a chitinderivative derived from crustaceans wherein the chitin or a chitinderivative have been purified or modified.

8. The composition does not comprise chitin or a chitin derivative thatis 80% or more deacetylated. The composition does not comprise chitin ora chitin derivative that is 70%-100% deacetylated.

9. The composition does not comprise chitin or a chitin derivative thatis free of fungal, micro-fungal, and crustacean chitin or chitinderivatives.

10. The composition does not comprise one or more coagulants, forexample: alpha-2-antiplasmin, alpha-1-antitrypsin,alpha-2-macroglobulin, aminohexanoic acid, aprotinin, a source ofCalcium ions, calcium alginate, calcium-sodium alginate, casein kinaseII, chitin, chitosan, collagen, cyanoacrylates, epsilon-aminocaproicacid, Factor XIII, fibrin, fibrin glue, fibrinogen, fibronectin,gelatin, living platelets, metha crylates, PAI-1, PAI-2, plasminactivator inhibitor, plasminogen, platelet agonists, protamine sulfate,prothrombin, an RGD peptide, sphingosine, a sphingosine derivative,thrombin, thromboplastin, and/or tranexamic acid.

11. The composition does not comprise epinephrine.

In certain embodiments of the invention, the composition of theinvention comprises a vasoconstrictor and/or a coagulant with one ofmore of the above provisos in certain embodiments.

In one embodiment, compositions of the invention encompass purifiedvasoconstrictors and/or coagulants, which may be about 100%, 90%, 80%,70%, 60%, 50%, 40%, 30%, or 20% pure. In a related embodiment,compositions of the invention encompass purified vasoconstrictors and/orcoagulants, which range between 100% and 20% in purity. In a preferredembodiment, the vasoconstrictors and/or coagulants is 90-100% purified.In another embodiment, the composition can consist of approximately100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10% or less of avasoconstrictor or coagulant. In a related embodiment, the compositioncan consist of between 100%, and 10% or less of a vasoconstrictor orcoagulant. Thus, the use of a vasoconstrictor and/or coagulant withvarying degrees of purity is encompassed by the invention.

Vasoconstrictors and/or coagulants that function at the desired distancecan be identified using the methods described herein. For example, thecardiac catheterization procedure simulation in dogs described inExample 1, of Section 6 can be used to measure the effect of potentialvasoconstrictors and/or coagulants at varying distances. The onlyportion of the procedure that needs to actually be simulated is thepuncture of a vein or artery with a catheter, mechanical observation ormanipulation to observe the vasculature associated with the heart is notnecessary. The identification of vasoconstrictors and/or coagulants thatfunction at varying distances can be achieved by selecting veins orarteries at specific distances beneath the skin surface for catheterpuncture. The distance of such veins or arteries can be determined byimaging techniques such as those disclosed in Section 5.2.

The composition can be formulated as a gel, solid, liquid, sponge, foam,spray, emulsion, suspension, solution, string, microbead, microsphere,or microfibril. The pharmaceutical compositions of the invention caninclude a pharmaceutically acceptable carrier, a neutral liquid, neutralgel or neutral solid. In certain preferred embodiments, wherein thecomposition has been formulated as a barrier, membrane, or film or thecomposition has been added to a barrier, membrane, or film, the neutralsolid or solid composition can be provided as a barrier, membrane, orfilm. A barrier, membrane, or film can be supplied in a variety ofstandard sizes, which can be further cut and sized to the area beingtreated. The barrier, membrane, or film can be a conventional bandage orgauze to which the composition of the invention is added or coated on,prior to application to the patient. Alternatively, the composition canbe formulated as a barrier, membrane, or film made out of strings,microbeads, microspheres, or microfibrils, or the composition can beformulated as a barrier-forming mat.

The pharmaceutical compositions of the invention can include a backing.For example, if the composition is formulated as a patch, a backing canbe adhered to the patch. The backing can be coated or embedded with anyadhesive compound so that areas of the backing that contact the skinwill adhere the backing and the attached composition of the invention tothe skin surface of the patient. The type of adhesive used can be anytype of medically acceptable adhesive. Such backings can be made ofnatural polymers or synthetic materials. Natural polymers from which thebacking can be made include but are not limited to cellulose and xylan.Synthetic materials from which the backing can be made include but arenot limited to polyurethane, Teflon, Dacron, stainless steel meshscreen, and a polyester woven fabric. Preferably the backing andadhesive are porous to areas which contact the skin to allow diffusionof oxygen. The backing can also serve as a surface upon which manualcompression can be applied.

The composition may be formulated as a barrier-forming material thatforms a barrier to blood. The composition can coat, be added to, orintegrated into a barrier-forming material that forms a barrier toblood. In one embodiment, the pharmaceutical composition comprises apatch made of barrier-forming materials that are embedded with one ormore coagulant and/or vasoconstrictor agent(s). In one embodiment, thepharmaceutical composition comprises a gauze embedded with one or morecoagulant and/or vasoconstrictor agent(s). In certain embodiments, thepharmaceutical compositions comprise a barrier-forming material embeddedor combined with one or more coagulant and/or vasoconstrictor agent(s),wherein the barrier-forming material contains an adhesive so that thematerial can be adhere to a patient's skin surface. Alternatively, thecomposition can lack barrier-forming materials.

According to one aspect of the invention, where the composition isformulated as or applied or integrated in a patch, the patch may vary insize. For example, the patch may be about 2 cm² to about 30 cm² in area.In certain embodiments, the patch is round, square, or rectangular. Inpreferred embodiments, the patch is 2 cm×2 cm, 3 cm×3 cm, 4 cm×4 cm, 5cm×5 cm, or 6 cm×6 cm in size. In other preferred embodiments, the patchcan be cut to reduce the size or to create a two dimensional shape asneeded. In preferred embodiments, the above described patches can bemanufactured for use in the methods, compositions, and kits of theinvention for adult patients weighing about 60, 70, 80, or 90 kg.

In related embodiments, the composition of the invention can containcollagen that can act as a sterile carrier.

In a preferred embodiment, the composition may include one or morecoagulants. The coagulant(s), for example, can be one or more of thefollowing: alpha-2-antiplasmin, alpha-1-antitrypsin,alpha-2-macroglobulin, aminohexanoic acid, aprotinin, a source ofCalcium ions, calcium alginate, calcium-sodium alginate, casein kinaseII, chitin, chitosan, collagen, cyanoacrylates, epsilon-aminocaproicacid, Factor XIII, fibrin, fibrin glue, fibrinogen, fibronectin,gelatin, living platelets, metha crylates, PAI-1, PAI-2, plasminactivator inhibitor, plasminogen, platelet agonists, protamine sulfate,prothrombin, an RGD peptide, sphingosine, a sphingosine derivative,thrombin, thromboplastin, and tranexamic acid.

In related embodiments, the composition of the invention can contain ananti-fungal or antibacterial agent, to prevent infection of the woundsurrounding the breach or puncture in the vein or artery.

In a preferred embodiment, the composition may include one or morevasoconstrictors. The vasoconstrictor(s), for example, can be one ormore of the following: endothelin, endothelin-1, epinephrine,adrenaline, metaraminol bitartrate (for example, manufactured asAramine™), dopamine HCl (for example, manufactured as Intropine™),isoproterenol HCl (for example, manufactured as Isuprel™),norepinephrine (for example, manufactured as Levophed™), phenylephrine,serotonin, thromboxane, norepinephrine, prostaglandin, methergine,oxytocin, isopreland U-46619, papaverine, yohimbine, visnadin, khellin,bebellin, and nicotinate derivatives.

In certain embodiments, the composition comprises a catalytic surfacethat accelerates a rate of a chemical reaction between the compositionof the invention and endogenous agents involved in hemostasis. Thecatalytic surface decreases the time to hemostasis and decreases thetime to achieve cessation or reduction of blood flow from the breach orpuncture in the vein or artery. The catalytic surface binds bloodcomponents to form a plug. The catalytic surface comprises a catalystthat is active at a distance, a distance as described herein in thecontext of the invention. The catalyst can reduce hemostasis time bycausing rapid release of vasoconstrictors substances and/or increasingthe concentration of vasoconstrictor substances. In other embodiments,the composition comprises a catalytic surface that accelerates a rate ofa chemical reaction between endogenous agents involved in hemostasis,such as the conversion of fibrinogen to fibrin. In yet otherembodiments, the composition comprises a catalytic surface thataccelerates or inhibits endogenous processes involved in hemostasis,such as release of serotonin by platelets or release of prothrombin orchemical signals by (imaged tissues. The chemical reaction or cellularprocess may be involved with vascular constriction that normally followsinjury to a blood vessel such as platelet plug formation, i.e. adherenceof platelets to the exposed basement membrane and connective tissue,change in shape of platelets from their discoid shape, secretion of ADP,secretion of serotonin, secretion of phospholipid, synthesis of plateletfactors, activation of phospholipase C, liberation of Calcium ions,activation of phospholipase A2, liberation of arachidonic acid,secretion of plasma clotting factors. The compositions of the inventioncan increase nucleation and formation of clots and/or the phasetransition of blood components from liquid to solid during the formationof a clot.

In one embodiment, the catalytic surface comprises fibers that areundissolveable and which are woven into a mat.

In certain embodiments, the compositions of the invention comprisevasoconstrictors and/or coagulants that are evenly distributedthroughout the composition, or supplied as a gradient, for example, bycombining fibrous pulps containing a range of desired concentrations ofvasoconstrictors and/or coagulants to produce a multilayeredcomposition.

The composition of the invention can contain a biodegradable material.Examples of biodegradable materials which can be used in the compositionof the invention include polyanionic polysaccharides, alginic acid,collagen, polyglycolide, polylactide, polycaprolactone, dextran andcopolymers thereof, polyglycolide, polylactide, polydioxanones,polyestercarbonates, polyhydroxyalkonates, polycaprolactone, andcopolymers thereof. Such biodegradable materials may form abarrier-forming portion of the composition, or act as carriers.

The pharmaceutical compositions of the invention can also comprisewound-healing and/or pain-reducing agents. Such agents includeanti-inflamatory agents, both steroidal and non-steroidal, such as butnot limited to agents which inhibit leukocyte migration into the area ofthe breach or puncture in the blood vessel (i.e., silver sulfadiazinemacetylsalicylic acid, indomethacin, and Nafazatrom), anti-histamines(i.e., pyrilamine, chlorpheniramine, tetraydrozoline, antazoline,cortisone, hydrocortisone, beta-metbasone, dexamethasone, fluocortolone,prednisolone, triamcinolone, indomethacin, and sulindac, its salts andits corresponding sulfide); agents which inhibit free radical formation(i.e., superoxide dismutase (SOD), catalase, glutathione peroxidase,β-carotene, ascorbic acid, transferring, ferritin, ceruloplasmin, anddesferrioxamine alpha-tocophenol); and bacteriostatic or bacteriocidalagents (i.e., cefoxitin, n-formamidoyl thienamycin, tetra cyclines,chloramphenicol, neomycin, gramicidin, bacitracin, sulfonamides,gentamycin, kanamycin, amikacin, sisomicin, tobramycin, norfloxican,nitrofurazones, and the combination of fluoroalanin/pentizidone).

5.4 Effective Dose

Generally, a therapeutically effective amount of a vasoconstrictorand/or coagulant for use in the methods, compositions, and kits of theinvention, will vary with the patients age, condition, and sex, as wellas the nature and extent of the condition in the subject, all of whichcan be determined by one of ordinary skill in the art. For example, theeffective dose needed for an infant may differ from an elderly patient,due to the reduced elasticity of blood vessels that accompanies age. Theeffective dose can also be dependent on the depth beneath the skinsurface at which a breach or puncture in a blood vessel is beingtreated. The dosage of the vasoconstrictor and/or coagulant of theinvention can be adjusted to accommodate the particular subject andcondition being treated.

In certain embodiments, the effective amount of the vasoconstrictorand/or coagulant does not directly increase or decrease the function ofcells involved in wound healing such as but not limited topolymorphonuclear leukocytes, macrophages and fibroblasts.

Toxicity and efficacy of the vasoconstrictor and/or coagulant of theinvention can be determined by standard pharmaceutical procedures incell cultures or experimental animals, e.g., for determining the ED50(the dose therapeutically effective in 50% of the population).Vasoconstrictors and/or coagulants that exhibit greater therapeuticeffect are preferred. In the present instance, vasoconstrictors and/orcoagulants that exhibit toxic side effects may be used in carrying outthe methods of the invention. The potential damage to unaffected cellsis minimized, since the vasoconstrictors and/or coagulants are appliedto the site of affected tissue and thereby reduce the risk of sideeffects.

The data obtained from the cell culture assays and animal studies can beused in formulating a range of dosage for use in humans. The dosage ofsuch vasoconstrictors and/or coagulants lies preferably within a rangeof concentrations that include the ED50. The dosage may vary within thisrange depending upon the formulation of the composition, i.e., gel,foam, patch, etc. For any vasoconstrictor used in the method of theinvention, the therapeutically effective dose can be estimated initiallyfrom tissue or tissue culture assays.

One standard tissue assay is conducted using aortic rings excised fromrats. The aorta are then rapidly suspended in a warmed Krebs-Henseleit(KH) buffer consisting of (in mmol/l): 118 NaCl, 4.75 KCl, 2.54CaCI₂.2H₂O, 1.19 KH₂PO₄, 1.19 MgSO₄.7H₂O, 12.5 NaHCO₃, and 10.0 glucose.Isolated vessels can be carefully freed of connective tissue and cutinto rings 2-3 mm in length. The rings are then mounted on stainlesssteel hooks, suspended in a 10-ml tissue bath, and connected to FT-03force displacement transducers (Grass Instrument, Quincy, Mass.) torecord changes in force on a Grass model 7 oscillographic recorder. Thebaths are filled with KH buffer and aerated at 37° C. with 95% O₂+5%CO₂. A resting force of 0.5 g is applied to the SMA rings, and then therings are equilibrated for 90 minutes. During this period, the buffer inthe tissue bath is replaced every 15-20 minutes, and the resting forceof the vascular rings is adjusted until 0.5 g of pre-load is maintained.After 90 to 120 minutes of equilibration, the rings are exposed to 100nM U-46619 (9,11-dideoxy-9α-11α-methaneepoxy-prostagalandin F_(2 α)Biomol Research Laboratories, Plymouth Meeting, Pa.), a thromboxane A₂mimetic, to generate 1.0 g of developed force. Once a stable contractionis obtained, acetylcholine, a typical endothelium-dependent vasodilator,is added to the bath in cumulative concentrations of 0.1, 1, 10, and 100nM to assess the integrity of endothelium. After the cumulative responseis stabilized, the rings are washed and again allowed to equilibrate tobaseline.

This procedure can be used with a vasoconstrictor other than U-46619 todetermine the effectiveness of a vasoconstrictor at maintainingvasoconstriction. The procedure can be repeated with varyingconcentration of a vasoconstrictor to determine effective dosage.

The function of a coagulant can be tested by standard assays. In suchassays, normal human blood, without anticoagulant, is drawn and placedin several test tubes. The normal blood, without a composition of theinvention, is allowed to clot (usually within 10 minutes). Other samplesof normal blood are drawn and one milliliter aliquots are placed in testtubes with descending aliquots of a particular coagulant used in themethods, compositions, and kits of the invention for which one desiresto test coagulant properties. With such an assay one can readilydetermine how many tenths of a milliliter of the coagulant are effectiveat causing clotting at a greater rate or in less time than blood withouta coagulant. Variations on this standard assay can be conducted wherethe patient has had an anticoagulant introduced into the bloodstreamprior to withdrawal of blood. The results can be used to identifycoagulant for use in the methods, compositions, and kits of theinvention that can accelerate hemostasis, i.e. cessation of flow ofblood from a breach or puncture in a vein or artery.

In various embodiments, an amount of vasoconstrictor and/or coagulantused in the methods, compositions, and kits of the invention is anamount that is 0.5-fold, 0.75-fold, 1-fold, 2-fold, 3-fold, 4-fold,5-fold, 10-fold, 12-fold, 15-fold, 20-fold, 50-fold or 100-fold theeffective dosage, for example, the dosage determined to be effective intreating a breach or puncture in a vein or artery when applied at adistance from the breach or puncture.

The vasoconstrictors and/or coagulants can be formulated into solid,liquid or gel-type formulations for topical application. Theconcentration of vasoconstrictor and/or coagulants in solid, liquidand/or gel-type formulations can be extrapolated from the dosage and thevolume or surface area of the formulation to be applied. For example,such compositions can be manufactured for general use for adults, e.g.,adults weighing about 50, 60, 70, 80, or 90 kg, and a topicaladministration area, e.g., of a patch, of about 1-2, 2-4, 4-8, 8-12,12-15, 15-20, 20-25, or 25-30 cm², or a topical administration volume,e.g., of a liquid or gel, of about 0.25-0.5, 0.5-1, 1-1.25, 1.25-1.5,1.5-1.75, 1.75-2, 2-2.5, or 2.5-3 ml. Exemplary concentrationcalculations are provided in Section 5.4.1, infra.

In embodiments of the invention where the composition of the inventionis formulated as, embedded in, or applied to a patch, 100 mg of thevasoconstrictor and/or coagulant may be present in 1 cm² of thewound-contacting surface of the patch. In other embodiments, theeffective amount of a vasoconstrictor and/or coagulant for use in themethods, compositions, and kits of the invention present in 1 cm² of apatch can be about 0.05 mg, 0.10 mg, 0.25 mg, 0.50 mg, 0.75 mg, 1 mg, 2mg, 5 mg, 8 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg,50 mg, 55 mg, 60 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 100 mg,150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg,600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 1000 mg, or 2000mg. In a preferred embodiment, the effective amount of a vasoconstrictorand/or coagulant of the invention present in 1 cm² of a patch is between0.05 mg and 30 mg of the vasoconstrictor and/or coagulant.

In other embodiments, where the vasoconstrictor and/or coagulant of theinvention is formulated as, embedded in, or applied to a patch, 100 μgof the vasoconstrictor and/or coagulant may be present in 1 cm² of thewound-contacting surface of the patch. In other embodiments, theeffective amount of a vasoconstrictor and/or coagulant of the inventionpresent in 1 cm² of a patch can be about 5 μg, 10 μg, 20 μg, 25 μg, 30μg, 35 μg, 40 μg, 45 μg, 50 μg, 55 μg, 60 μg, 65 μg, 70 μg, 75 μg, 80μg, 85 μg, 90 μg, 95 μg, 100 μg, 105 μg, 110 μg, 115 μg, 120 μg, 125 μg,130 μg, 135 μg, 140 μg, 145 μg, 150 μg, 155 μg, or 160 μg between about1 mM and 70 mM. An example of a coagulant that can be effective in suchamounts is thrombin. An example of a vasoconstrictor that can beeffective in such amounts is endothelin-1.

In preferred embodiments of the present methods and compositions, thedose of a vasoconstrictor and/or coagulant, e.g., on the surface of apatch, regardless of its molecular weight, is about 1 mM to about 70 mM.In certain exemplary embodiments, the dose is about 1 mM to about 10 mM,about 10 mM to about 30 mM, about 30 mM to about 50 mM, or about 50 mMto about 70 mM.

In yet other embodiments, the effective amount of a vasoconstrictorand/or coagulant of the invention is about 1-1000 IU/cm² wherein thevasoconstrictor and/or coagulant is formulated as, embedded in, orapplied to a patch.

In one embodiment, the vasoconstrictor and/or coagulant forms aconcentration gradient that decreases from the site of application tothe breach or puncture in the vein or artery. For example, in the caseof a cardiac catheterization track wound, the vasoconstrictor and/orcoagulant can form a concentration gradient through the track wound andpromote clot formation throughout the track resulting in an decreased inthe time necessary to achieve hemostasis.

In certain embodiments, an effective amount of a vasoconstrictor and/orcoagulant of the invention is an amount that activates hemostasis in thepresence of a coagulant or an anticoagulant.

In certain embodiments, the effective dose is the dose necessary toinitiate clotting with or without compression. In other embodiments, theeffective dose is the dose necessary to cause formation of a firm clotthat will remain with or without compression. In yet other embodiments,an effective dose can be determined by the strength of the clot, i.e.the time for which the clot holds with or without compression.

Once it has been determined how varying concentrations and amounts of aparticular vasoconstrictor and/or coagulant act in vitro, effectivevasoconstrictors and/or coagulants can be further tested in animalmodels for the distance at which they function by methods described inSection 5.5. An effective amount of a vasoconstrictor and/or coagulantthat functions at a given distance can be determined by measuring thetime to form a clot and the strength of a clot sealing a breach orpuncture in a vein or artery with thromboelastography. A series ofmeasurements can be taken varying the concentration or amount of thevasoconstrictor and/or coagulant to determine an effective amount. Thedistance of a breach or puncture in a vein or artery from the skinsurface can also be varied to determine the maximum or optimal effectivedistance. Such series of measurements can be used to predict how aparticular vasoconstrictor and/or coagulant will function at aparticular distance, and allow for a determination of effective amountfor a desired distance. The distance animal veins or arteries arebeneath the skin surface can be determined by imaging techniques such asthose disclosed in Section 5.4. The distance at which certainconcentrations or amounts of vasoconstrictors and/or coagulants areeffective in vivo can be used to extrapolate the effective amount for adesired distance needed for a particular patient, or the limit to thedistance at which a particular vasoconstrictor and/or coagulant iseffective. Such information can be used to more accurately determineuseful doses in humans.

Results from animal models can be extrapolated to determine effectivedoses for human subjects. Comparing varying concentrations of avasoconstrictor and/or coagulant in one or more animal models allows forthe establishment of dose response curves that can be used to estimateeffective amounts in a human, given the particular circumstances of eachsubject, i.e. distance of wound, size of wound, presence of coagulantsor anticoagulants in the blood stream.

Human patients can also be used to determine the distance at which avasoconstrictor and/or coagulant is effective. For example, ultrasoundcan be used to visualize the distance a breach or puncture in a vein orartery is from the surface of the skin and determine if blood flow fromthe breach or puncture has decreased or been eliminated. Ultrasoundprobes can be used to locate the breached or punctured vein or artery orto image a specific area by aligning the probes with a desired vein orartery. Ultrasound imaging of the breach or puncture site can becombined with Doppler flow analysis. Doppler flow analysis allows forthe determination of cessation or reduction of blood flow through theartery or vein. If blood flow through the vein or artery is inhibited,it may cause damage. Thus, combining Doppler flow and ultrasound wouldallow for a determination of the maximum upper limit of an effectiveamount of a vasoconstrictor and/or coagulant. For example, if the effectof a coagulant extends into the blood vessel and causes clotting ofplatelet and cessation of blood within the blood vessel, the effectcould be damaging. In another embodiment, the maximum upper limit of aneffective amount of a vasoconstrictor and/or coagulant can be measuredas the amount of the vasoconstrictor and/or coagulant for use in themethods, compositions, and kits of the invention that causes an increasein cardiac output.

5.4.1 Vasoconstrictor Doses

In certain preferred embodiments, the effective amount of avasoconstrictor used in the compositions of the methods, compositions,and kits of the invention will vary with the patients age, condition,and sex, as well as the nature and extent of the condition in thesubject, all of which can be determined by one of ordinary skill in theart. For example, a patients body weight can be used to determine aneffective amount of a vasoconstrictor.

In one embodiment, the composition topically administered to a patientin the methods of the invention comprises an effective amount of thevasoconstrictor Adrenaline™ in a concentration in the range of about0.000001 mg/kg of patient body weight to about 11 mg/kg of patient bodyweight. For adults the preferred Adrenaline™ concentration range isabout 0.00001 mg/kg to about 0.5 mg/kg. In related embodiments theeffective amount of Adrenaline™ is approximately 0.00001 mg/kg, 0.00005mg/kg, 0.0001 mg/kg, 0.0005 mg/kg, 0.001 mg/kg, 0.005 mg/kg, 0.01 mg/kg,0.05 mg/kg, 0.1 mg/kg, 0.5 mg/kg, 1.0 mg/kg, 1.5 mg/kg, 2.0 mg/kg, 2.5mg/kg, 3.0 mg/kg, 3.5 mg/kg, 4.0 mg/kg, 4.5 mg/kg, 5.0 mg/kg, 5.5 mg/kg,6.0 mg/kg, 6.5 mg/kg, 7.0 mg/kg, 7.5 mg/kg, 8.0 mg/kg, 8.5 mg/kg, 9.0mg/kg, 9.5 mg/kg, 10.0 mg/kg, 10.5 mg/kg, or 11.0 mg/kg of patient bodyweight. In related embodiments the effective amount of Adrenaline™ isabout 0.00001 mg/kg to 0.0001 mg/kg, 0.001 mg/kg to 0.01 mg/kg, 0.1mg/kg to 1.0 mg/kg, 2.0 mg/kg to 3.0 mg/kg, 4.0 mg/kg to 5.0 mg/kg, 6.0mg/kg to 7.0 mg/kg, 8.0 mg/kg to 9.0 mg/kg, or 10.0 mg/kg to 11.0 mg/kgof patient body weight. In related embodiments the effective amount ofAdrenaline™ is about 0.00005 mg/kg to 0.0005 mg/kg, 0.005 mg/kg to 0.05mg/kg, 0.5 mg/kg to 1.5 mg/kg, 2.5 mg/kg to 3.5 mg/kg, 4.5 mg/kg to 5.5mg/kg, 6.5 mg/kg to 7.5 mg/kg, 8.5 mg/kg to 9.5 mg/kg, or 10.5 mg/kg to11.5 mg/kg of patient body weight

In another embodiment, the composition topically administered to apatient in the methods of the invention comprises an effective amount ofthe vasoconstrictor metaraminol bitartrate in a concentration in therange of about 0.00001 mg/kg of patient body weight to about 1500 mg/kgof patient body weight. For adults the preferred metaraminol bitartrateconcentration range is about 0.0005 mg/kg to about 4.5 mg/kg. In relatedembodiments the effective amount of metaraminol bitartrate isapproximately 0.0005 mg/kg, 0.001 mg/kg, 0.005 mg/kg, 0.01 mg/kg, 0.05mg/kg, 0.1 mg/kg, 0.5 mg/kg, 1.0 mg/kg, 10 mg/kg, 50 mg/kg, 100 mg/kg,150 mg/kg, 200 mg/kg, 300 mg/kg, 400 mg/kg, 500 mg/kg, 600 mg/kg, 700mg/kg, 800 mg/kg, 900 mg/kg, 1000 mg/kg, 1100 mg/kg, 1200 mg/kg, 1300mg/kg, 1400 mg/kg, or 1500 mg/kg of patient body weight. In relatedembodiments the effective amount of metaraminol bitartrate is about0.0005 mg/kg to 0.005 mg/kg, 0.05 mg/kg to 0.5 mg/kg, 50 mg/kg to 150mg/kg, 300 mg/kg to 500 mg/kg, 700 mg/kg to 900 mg/kg, 1100 mg/kg to1300 mg/kg, or 1300 mg/kg to 1500 mg/kg of patient body weight. Inrelated embodiments the effective amount of metaraminol bitartrate isabout 0.001 mg/kg to 0.01 mg/kg, 0.1 mg/kg to 1.0 mg/kg, 10 mg/kg to 100mg/kg, 200 mg/kg to 400 mg/kg, 600 mg/kg to 800 mg/kg, 1000 mg/kg to1200 mg/kg, or 1200 mg/kg to 1400 mg/kg of patient body weight.

In one embodiment, the composition topically administered to a patientin the methods of the invention comprises an effective amount of thevasoconstrictor dopamine HCl in a concentration in the range of about0.00001 mg/kg of patient body weight to about 150 mg/kg of patient bodyweight. For adults the preferred dopamine HCl concentration range isabout 0.0005 mg/kg to about 10 mg/kg. In related embodiments theeffective amount of dopamine HCl is approximately 0.0005 mg/kg, 0.001mg/kg, 0.005 mg/kg, 0.01 mg/kg, 0.05 mg/kg, 0.1 mg/kg, 0.5 mg/kg, 1.0mg/kg, 10 mg/kg, 20 mg/kg, 30 mg/kg, 40 mg/kg, 50 mg/kg, 60 mg/kg, 70mg/kg, 80 mg/kg, 90 mg/kg, 100 mg/kg, 110 mg/kg, 120 mg/kg, 130 mg/kg,140 mg/kg, or 150 mg/kg of patient body weight. In related embodimentsthe effective amount of dopamine HCl is about 0.0005 mg/kg to 0.005mg/kg, 0.05 mg/kg to 0.5 mg/kg, 10 mg/kg to 30 mg/kg, 50 mg/kg to 70mg/kg, 90 mg/kg to 110 mg/kg, or 130 mg/kg to 150 mg/kg of patient bodyweight. In related embodiments the effective amount of dopamine HCl isabout 0.001 mg/kg to 0.01 mg/kg, 0.1 mg/kg to 1.0 mg/kg, 10 mg/kg to 20mg/kg, 40 mg/kg to 60 mg/kg, 80 mg/kg to 100 mg/kg, or 120 mg/kg to 140mg/kg of patient body weight.

In one embodiment, the composition topically administered to a patientin the methods of the invention comprises an effective amount of thevasoconstrictor isoproterenol HCl in a concentration in the range ofabout 0.00001 mg/kg of patient body weight to about 150 mg/kg of patientbody weight. For adults the preferred isoproterenol HCl concentrationrange is about 0.0005 mg/kg to about 5 mg/kg. In related embodiments theeffective amount of isoproterenol HCl is approximately 0.0001 mg/kg,0.0005 mg/kg, 0.001 mg/kg, 0.005 mg/kg, 0.01 mg/kg, 0.05 mg/kg, 0.1mg/kg, 0.5 mg/kg, 1.0 mg/kg, 10 mg/kg, 20 mg/kg, 30 mg/kg, 40 mg/kg, 50mg/kg, 60 mg/kg, 70 mg/kg, 80 mg/kg, 90 mg/kg, 100 mg/kg, 110 mg/kg, 120mg/kg, 130 mg/kg, 140 mg/kg, or 150 mg/kg of patient body weight. Inrelated embodiments the effective amount of isoproterenol HCl is about0.0001 mg/kg to 0.001 mg/kg, 0.01 mg/kg to 0.1 mg/kg, 1.0 mg/kg to 20mg/kg, 40 mg/kg to 60 mg/kg, 80 mg/kg to 100 mg/kg, or 120 mg/kg to 140mg/kg of patient body weight. In related embodiments the effectiveamount of isoproterenol HCl is about 0.0005 mg/kg to 0.005 mg/kg, 0.05mg/kg to 0.1 mg/kg, 0.5 mg/kg to 10 mg/kg, 30 mg/kg to 50 mg/kg, 70mg/kg to 90 mg/kg, 110 mg/kg to 130 mg/kg, or 130 mg/kg to 150 mg/kg ofpatient body weight.

In one embodiment, the composition topically administered to a patientin the methods of the invention comprises an effective amount of thevasoconstrictor norepinephrine in a concentration in the range of about0.00001 mg/kg of patient body weight to about 2 mg/kg of patient bodyweight. For adults the preferred norepinephrine concentration range isabout 0.0001 mg/kg to about 0.01 mg/kg. In related embodiments theeffective amount of norepinephrine is approximately 0.0001 mg/kg, 0.0005mg/kg, 0.001 mg/kg, 0.005 mg/kg, 0.01 mg/kg, 0.05 mg/kg, 0.1 mg/kg, 0.5mg/kg, 1.0 mg/kg, 1.5 mg/kg, or 2 mg/kg of patient body weight. Inrelated embodiments the effective amount of norepinephrine is about0.0001 mg/kg to 0.001 mg/kg, 0.01 mg/kg to 0.1 mg/kg, or 1.0 mg/kg to 2mg/kg of patient body weight. In related embodiments the effectiveamount of norepinephrine is about 0.0005 mg/kg to 0.005 mg/kg, 0.05mg/kg to 0.5 mg/kg, or 1.5 mg/kg to 2 mg/kg of patient body weight.

In one embodiment, the composition topically administered to a patientin the methods of the invention comprises an effective amount of thevasoconstrictor Serotonin™ in a concentration in the range of about0.0001 mg/kg of patient body weight to about 750 mg/kg of patient bodyweight. For adults the preferred Serotonin™ concentration range is about0.001 mg/kg to about 0.6 mg/kg. In related embodiments the effectiveamount of Serotonin™ is approximately 0.001 mg/kg, 0.005 mg/kg, 0.01mg/kg, 0.05 mg/kg, 0.1 mg/kg, 0.5 mg/kg, 1.0 mg/kg, 10 mg/kg, 20 mg/kg,30 mg/kg, 40 mg/kg, 50 mg/kg, 60 mg/kg, 70 mg/kg, 80 mg/kg, 90 mg/kg,100 mg/kg, 110 mg/kg, 120 mg/kg, 130 mg/kg, 140 mg/kg, 150 mg/kg, 160mg/kg, 170 mg/kg, 180 mg/kg, 190 mg/kg, 200 mg/kg, 210 mg/kg, 250 mg/kg,300 mg/kg, 350 mg/kg, 400 mg/kg, 450 mg/kg, 500 mg/kg, 550 mg/kg, 600mg/kg, 650 mg/kg, 700 mg/kg, or 750 mg/kg of patient body weight. Inrelated embodiments the effective amount of Serotonin™ is about 0.001mg/kg to 0.01 mg/kg, 0.1 mg/kg to 1.0 mg/kg, 20 mg/kg to 40 mg/kg, 60mg/kg to 80 mg/kg, 100 mg/kg to 120 mg/kg, 140 mg/kg to 160 mg/kg, 180mg/kg to 200 mg/kg, 210 mg/kg to 300 mg/kg, 400 mg/kg to 500 mg/kg, or600 mg/kg to 700 mg/kg of patient body weight. In related embodimentsthe effective amount of Serotonin™ is about 0.005 mg/kg to 0.05 mg/kg,0.5 mg/kg to 10 mg/kg, 30 mg/kg to 50 mg/kg, 70 mg/kg to 90 mg/kg, 110mg/kg to 130 mg/kg, 150 mg/kg to 170 mg/kg, 190 mg/kg to 210 mg/kg, 250mg/kg to 350 mg/kg, 450 mg/kg to 550 mg/kg, or 650 mg/kg to 750 mg/kg ofpatient body weight.

In one embodiment, the composition topically administered to a patientin the methods of the invention comprises an effective amount of thevasoconstrictor endothelin in a concentration in the range of about4.5×10⁻⁸ mg/kg of patient body weight to about 5.0×10⁻⁶ mg/kg of patientbody weight. In related embodiments the effective amount of endothelinis approximately 4.5×10⁻⁸ mg/kg, 5.0×10⁻⁸ mg/kg, 5.5×10⁻⁸ mg/kg,6.0×10⁻⁸ mg/kg, 6.5×10⁻⁸ mg/kg, 7.0×10⁻⁸ mg/kg, 7.5×10⁻⁸ mg/kg, 8.0×10⁻⁸mg/kg, 8.5×10⁻⁸ mg/kg, 9.0×10⁻⁸ mg/kg, 9.5×10⁻⁸ mg/kg, 1.0×10⁻⁷ mg/kg,1.5×10⁻⁷ mg/kg, 2.0×10⁻⁷ mg/kg, 2.5×10⁻⁷ mg/kg, 3.0×10⁻⁷ mg/kg, 3.5×10⁻⁷mg/kg, 4.0×10⁻⁷ mg/kg, 4.5×10⁻⁷ mg/kg, 5.0×10⁻⁷ mg/kg, 5.5×10⁻⁷ mg/kg,6.0×10⁻⁷ mg/kg, 6.5×10⁻⁷ mg/kg, 7.0×10⁻⁷ mg/kg, 7.5×10⁻⁷ mg/kg, 8.0×10⁻⁷mg/kg, 8.5×10⁻⁷ mg/kg, 9.0×10⁻⁷ mg/kg, 1.0×10⁻⁶ mg/kg, 1.5×10⁻⁶ mg/kg,2.0×10⁻⁶ mg/kg, 2.5×10⁻⁶ mg/kg, 3.0×10⁻⁶ mg/kg, 3.5×10⁻⁶ mg/kg, 4.0×10⁻⁶mg/kg, 4.5×10⁻⁶ mg/kg, or 5.0×10⁻⁶ mg/kg of patient body weight. Inrelated embodiments the effective amount of endothelin is about 5.0×10⁻⁸mg/kg to 6.0×10⁻⁸ mg/kg, 7.0×10⁻⁸ mg/kg to 8.0×10⁻⁸ mg/kg, 9.0×10⁻⁸mg/kg to 1.0×10⁻⁷ mg/kg, 2.0×10⁻⁷ mg/kg to 3.0×10⁻⁷ mg/kg, 4.0×10⁻⁷mg/kg to 5.0×10⁻⁷ mg/kg, 6.0×10⁻⁷ mg/kg to 7.0×10⁻⁷ mg/kg, 8.0×10⁻⁷mg/kg to 9.0×10⁻⁷ mg/kg, 1.0×10⁻⁶ mg/kg to 2.0×10⁻⁶ mg/kg, 3.0×10⁻⁶mg/kg to 4.0×10⁻⁶ mg/kg, or 4.0×10⁻⁶ mg/kg to 5.0×10⁻⁶ mg/kg of patientbody weight. In related embodiments the effective amount of endothelinis about 4.5×10⁻⁸ mg/kg to 5.5×10⁻⁸ mg/kg, 6.5×10⁻⁸ mg/kg to 7.5×10⁻⁸mg/kg, 8.5×10⁻⁸ mg/kg to 9.5×10⁻⁸ mg/kg, 1.5×10⁻⁷ mg/kg to 2.5×10⁻⁷mg/kg, 3.5×10⁻⁷ mg/kg to 4.5×10⁻⁷ mg/kg, 5.5×10⁻⁷ mg/kg to 6.5×10⁻⁷mg/kg, 7.5×10⁻⁷ mg/kg to 8.5×10⁻⁷ mg/kg, 1.5×10⁻⁶ mg/kg to 2.5×10⁻⁶mg/kg, or 3.5×10⁻⁶ mg/kg to 4.5×10⁻⁶ mg/kg of patient body weight.

For other vasoconstrictors, including but not limited to, endothelin-1,epinephrine, phenylephrine, thromboxane, prostaglandin, methergine,oxytocin, isopreland U-46619, papaverine, yohimbine, visnadin, khellin,bebellin, and nicotinate derivatives, one skilled in the art would beable to determine appropriate effective doses using the methodsdescribed in sections 5.2 and 5.4. In related embodiments the effectiveamount of a vasoconstrictor is approximately 4.5×10⁻⁸ mg/kg, 5.0×10⁻⁸mg/kg, 6.0×10⁻⁸ mg/kg, 7.0×10⁻⁸ mg/kg, 8.0×10⁻⁸ mg/kg, 9.0×10⁻⁸ mg/kg,1.0×10⁻⁷ mg/kg, 2.0×10⁻⁷ mg/kg, 3.0×10⁻⁷ mg/kg, 4.0×10⁻⁷ mg/kg, 5.0×10⁻⁷mg/kg, 6.0×10⁻⁷ mg/kg, 7.0×10⁻⁷ mg/kg, 8.0×10⁻⁷ mg/kg, 9.0×10⁻⁷ mg/kg,1.0×10⁻⁶ mg/kg, 2.0×10⁻⁶ mg/kg, 3.0×10⁻⁶ mg/kg, 4.0×10⁻⁶ mg/kg, 5.0×10⁻⁶mg/kg, 6.0×10⁻⁶ mg/kg, 7.0×10⁻⁶ mg/kg, 8.0×10⁻⁶ mg/kg, 9.0×10⁻⁶ mg/kg,1.0×10⁻⁵ mg/kg, 2.0×10⁻⁵ mg/kg, 3.0×10⁻⁵ mg/kg, 4.0×10⁻⁵ mg/kg, 5.0×10⁻⁵mg/kg, 6.0×10⁻⁵ mg/kg, 7.0×10⁻⁵ mg/kg, 8.0×10⁻⁵ mg/kg, 9.0×10⁻⁵ mg/kg,1.0×10⁻⁴ mg/kg, 2.0×10⁻⁴ mg/kg, 3.0×10⁻⁴ mg/kg, 4.0×10⁻⁴ mg/kg, 5.0×10⁻⁴mg/kg, 6.0×10⁻⁴ mg/kg, 7.0×10⁻⁴ mg/kg, 8.0×10⁻⁴ mg/kg, 9.0×10⁻⁴ mg/kg,1.0×10⁻³ mg/kg, 2.0×10⁻³ mg/kg, 3.0×10⁻³ mg/kg, 4.0×10⁻³ mg/kg, 5.0×10⁻³mg/kg, 6.0×10⁻³ mg/kg, 7.0×10⁻³ mg/kg, 8.0×10⁻³ mg/kg, 9.0×10⁻³ mg/kg,1.0×10⁻² mg/kg, 2.0×10⁻² mg/kg, 3.0×10⁻² mg/kg, 4.0×10⁻² mg/kg, 5.0×10⁻²mg/kg, 6.0×10⁻² mg/kg, 7.0×10⁻² mg/kg, 8.0×10⁻² mg/kg, 9.0×10⁻² mg/kg,0.1 mg/kg, 0.5 mg/kg, 1.0 mg/kg, 10 mg/kg, 20 mg/kg, 30 mg/kg, 40 mg/kg,50 mg/kg, 60 mg/kg, 70 mg/kg, 80 mg/kg, 90 mg/kg, 100 mg/kg, 110 mg/kg,120 mg/kg, 130 mg/kg, 140 mg/kg, 150 mg/kg, 160 mg/kg, 170 mg/kg, 180mg/kg, 190 mg/kg, 200 mg/kg, 210 mg/kg, 250 mg/kg, 300 mg/kg, 350 mg/kg,400 mg/kg, 450 mg/kg, 500 mg/kg, 550 mg/kg, 600 mg/kg, 650 mg/kg, 700mg/kg, 750 mg/kg, 800 mg/kg, 850 mg/kg, 900 mg/kg, 950 mg/kg, 1000mg/kg, 1100 mg/kg, 1200 mg/kg, 1300 mg/kg, 1400 mg/kg, or 1500 mg/kg ofpatient body weight.

In certain embodiments, the effective amount of a vasoconstrictor is inthe range of about 6.0×10⁻⁸ mg/kg to 1.0×10⁻⁷ mg/kg, 1.0×10⁻⁷ mg/kg to4.0×10⁻⁷ mg/kg, 4.0×10⁻⁷ mg/kg to 8.0×10⁻⁷ mg/kg, 8.0×10⁻⁷ mg/kg to2.0×10⁻⁶ mg/kg, 2.0×10⁻⁶ mg/kg to 6.0×10⁻⁶ mg/kg, 6.0×10⁻⁶ mg/kg to1.0×10⁻⁵ mg/kg, 1.0×10⁻⁵ mg/kg to 4.0×10⁻⁵ mg/kg, 4.0×10⁻⁵ mg/kg to8.0×10⁻⁵ mg/kg, 8.0×10⁻⁵ mg/kg to 2.0×10⁻⁴ mg/kg, 2.0×10⁻⁴ mg/kg to6.0×10⁻⁴ mg/kg, 6.0×10⁻⁴ mg/kg to 1.0×10⁻³ mg/kg, 1.0×10⁻³ mg/kg to4.0×10⁻³ mg/kg, 4.0×10⁻³ mg/kg to 8.0×10⁻³ mg/kg, 8.0×10⁻³ mg/kg to2.0×10⁻² mg/kg, 2.0×10⁻² mg/kg to 6.0×10⁻² mg/kg, 6.0×10⁻² mg/kg to 0.1mg/kg, 0.1 mg/kg to 0.4 mg/kg, 0.4 mg/kg to 0.8 mg/kg, 0.8 mg/kg to 2.0mg/kg, 2.0 mg/kg to 6.0 mg/kg, 6.0 mg/kg to 10 mg/kg, 10 mg/kg to 40mg/kg, 40 mg/kg to 80 mg/kg, 80 mg/kg to 120 mg/kg, 120 mg/kg to 160mg/kg, 160 mg/kg to 200 mg/kg, 200 mg/kg to 240 mg/kg, 280 mg/kg to 320mg/kg, 320 mg/kg to 360 mg/kg, 360 mg/kg to 400 mg/kg, 400 mg/kg to 440mg/kg, 440 mg/kg to 480 mg/kg, 480 mg/kg to 520 mg/kg, 520 mg/kg to 560mg/kg, 560 mg/kg to 600 mg/kg, 600 mg/kg to 640 mg/kg, 640 mg/kg to 680mg/kg, 680 mg/kg to 720 mg/kg, 720 mg/kg to 760 mg/kg, 760 mg/kg to 800mg/kg, 800 mg/kg to 840 mg/kg, 840 mg/kg to 880 mg/kg, 880 mg/kg to 920mg/kg, 920 mg/kg to 960 mg/kg, 960 mg/kg to 1000 mg/kg, 1000 mg/kg to1040 mg/kg, 1040 mg/kg to 1080 mg/kg, 1080 mg/kg to 1120 mg/kg, 1120mg/kg to 1160 mg/kg, 1160 mg/kg to 1200 mg/kg, 1200 mg/kg to 1240 mg/kg,1240 mg/kg to 1280 mg/kg, 1280 mg/kg to 1320 mg/kg, 1320 mg/kg to 1360mg/kg, 1360 mg/kg to 1400 mg/kg, 1400 mg/kg to 1440 mg/kg, 1440 mg/kg to1480 mg/kg, or 1480 mg/kg to 1520 mg/kg of patient body weight.

In certain embodiments, the effective amount of a vasoconstrictor is inthe range of about 4.5×10⁻⁸ mg/kg to 8.0×10⁻⁸ mg/kg, 8.0×10⁻⁸ mg/kg to2.0×10⁻⁷ mg/kg, 2.0×10⁻⁷ mg/kg to 6.0×10⁻⁷ mg/kg, 6.0×10⁻⁷ mg/kg to1.0×10⁻⁶ mg/kg, 1.0×10⁻⁶ mg/kg to 4.0×10⁻⁶ mg/kg, 4.0×10⁻⁶ mg/kg to8.0×10⁻⁶ mg/kg, 8.0×10⁻⁶ mg/kg to 2.0×10⁻⁵ mg/kg, 2.0×10⁻⁵ mg/kg to6.0×10⁻⁵ mg/kg, 6.0×10⁻⁵ mg/kg to 1.0×10⁻⁴ mg/kg, 1.0×10⁻⁴ mg/kg to4.0×10⁻⁴ mg/kg, 4.0×10⁻⁴ mg/kg to 8.0×10⁻⁴ mg/kg, 8.0×10⁻⁴ mg/kg to2.0×10⁻³ mg/kg, 2.0×10⁻³ mg/kg to 6.0×10⁻³ mg/kg, 6.0×10⁻³ mg/kg to1.0×10⁻² mg/kg, 1.0×10⁻² mg/kg to 4.0×10⁻² mg/kg, 4.0×10⁻² mg/kg to8.0×10⁻² mg/kg, 8.0×10⁻² mg/kg to 0.2 mg/kg, 0.2 mg/kg to 0.6 mg/kg, 0.6mg/kg to 1.0 mg/kg, 1.0 mg/kg to 4.0 mg/kg, 4.0 mg/kg to 8.0 mg/kg, 8.0mg/kg to 12 mg/kg, 12 mg/kg to 16 mg/kg, 16 mg/kg to 20 mg/kg, 20 mg/kgto 60 mg/kg, 60 mg/kg to 100 mg/kg, 100 mg/kg to 140 mg/kg, 140 mg/kg to180 mg/kg, 180 mg/kg to 220 mg/kg, 220 mg/kg to 260 mg/kg, 260 mg/kg to300 mg/kg, 300 mg/kg to 340 mg/kg, 340 mg/kg to 380 mg/kg, 380 mg/kg to420 mg/kg, 420 mg/kg to 460 mg/kg, 460 mg/kg to 500 mg/kg, 500 mg/kg to540 mg/kg, 540 mg/kg to 580 mg/kg, 580 mg/kg to 620 mg/kg, 620 mg/kg to660 mg/kg, 660 mg/kg to 700 mg/kg, 700 mg/kg to 740 mg/kg, 740 mg/kg to780 mg/kg, 780 mg/kg to 820 mg/kg, 820 mg/kg to 860 mg/kg, 860 mg/kg to900 mg/kg, 900 mg/kg to 940 mg/kg, 940 mg/kg to 980 mg/kg, 980 mg/kg to1020 mg/kg, 1020 mg/kg to 1060 mg/kg, 1060 mg/kg to 1100 mg/kg, 1100mg/kg to 1140 mg/kg, 1140 mg/kg to 1180 mg/kg, 1180 mg/kg to 1220 mg/kg,1220 mg/kg to 1260 mg/kg, 1260 mg/kg to 1300 mg/kg, 1300 mg/kg to 1340mg/kg, 1340 mg/kg to 1380 mg/kg, 1380 mg/kg to 1420 mg/kg, 1420 mg/kg to1460 mg/kg, or 1460 mg/kg to 1500 mg/kg of patient body weight.

In general, the effective does of a vasoconstrictor used in thecompositions of the methods, compositions, and kits of the invention isa concentration that is less than an amount that would cause systemicvasoconstriction when administered intravenously and greater than anamount that would have only a localized effect when applied to the skinsurface rather than effecting a breach or puncture in a vein or artery.

The effective amount of a vasoconstrictor can also be described in thecontext of particular formulations of the compositions used in themethods, compositions, and kits of the invention. For example, where thecomposition is formulated as or applied to a patch for administrationfollowing a cardiac catheterization procedure, adrenaline is thevasoconstrictor used in the composition, and the patient is an adultweighing about 70 kg, then the patch should comprise adrenaline in theconcentration range of about 0.00075 mg/kg to about 37.5 mg/kg. Since apatch used in such procedures is typically about 4 cm² to 25 cm²,depending on the F size of the catheter used, one skilled in the artcould readily determine the concentration in units of mg of adrenalineper cm² of patch, e.g., about 0.00020 mg/cm² to about 1.5 mg/cm².Similarly, for example, where the composition is formulated as a gel orliquid, for administration following a cardiac catheterizationprocedure, adrenaline is the vasoconstrictor used in the composition,and the patient is an adult weighing about 70 kg, then the gel shouldcomprise adrenaline in the concentration range of about 0.00075 mg/kg toabout 37.5 mg/kg. Since a gel used in such procedures is typically about1-2 ml, one skilled in the art could readily determine the concentrationin units of mg of adrenaline per ml of gel, e.g., about 0.00075 mg/ml toabout 18.75 mg/ml.

The example conversion calculation described above can be performed toallow the extrapolation of suitable concentration ranges for topicalapplication any of the dosage ranges for any of the vasoconstrictorsand/or coagulants described herein above or any vasoconstrictors and/orcoagulant suitable for use in the methods, compositions, and kits of theinvention. In preferred embodiments, where the composition is formulatedas a patch, gel, or liquid, such compositions can be manufactured forgeneral use for adults, e.g., adults weighing about 50, 60, 70, 80, or90 kg, and a topical administration area, e.g., of a patch, of about1-2, 2-4, 4-8, 8-12, 12-15, 15-20, 20-25, or 25-30 cm², or a topicaladministration volume, e.g., of a liquid or gel, of about 0.25-0.5,0.5-1, 1-1.25, 1.25-1.5, 1.5-1.75, 1.75-2, 2-2.5, or 2.5-3 ml.

5.5 Kits

A kit is also provided which according to the invention comprises any ofthe above described embodiments. The kit can include the compositioncontained within a sealed, water proof, sterile package whichfacilitates removal of the composition without contamination. Materialsfrom which containers may be made include aluminum foil, plastic, oranother conventional material that is easily sterilized. The kit cancontain a single composition or multiple compositions, preferablywherein each is provided in a separate, waterproof, sterile package.

In another embodiment, a container having dual compartments is provided.A first compartment contains the composition, while the secondcompartment contains a topical barrier, membrane, or film according tothe invention. In field use, the barrier, membrane, or film can bereadily dipped into an opened first compartment ans subsequently appliedto the wound. The composition can be applied or added to a topicalbarrier, membrane, or film prior to being packaged and sterilized or thecomposition can be formulated as a barrier, membrane, or film. Inanother embodiment, the kit may not contain a barrier, membrane, or filmif the composition is not formulated as such.

According to one aspect of the invention, various specialized kits canbe provided. The kit can contain multiple compositions of the invention,wherein each is contained within a separate sealed sterile package orcontainer. The kit can contain in one or more containers, a an effectiveamount of a vasoconstrictor, wherein the vasoconstrictor does notcomprise a poly-β-1→4 N-acetylglucosamine polymer or derivative thereof,while in another separate container a pharmaceutically acceptablecarrier can be packaged. In a separate container the kit can contain aneutral liquid, neutral gel or neutral solid. The kit may also contain acoagulant in accord with the composition of the invention as describedherein.

The kit can comprise a composition that can be formulated as abarrier-forming material that forms a barrier to blood. The kit cancomprise a composition that can coat, be added to, or integrated into abarrier-forming material that forms a barrier to blood. In oneembodiment a kit comprises a pharmaceutical composition comprising apatch made of barrier-forming materials that are embedded with one ormore coagulant and/or vasoconstrictor agent(s). In one embodiment a kitcomprises a pharmaceutical composition comprising a gauze embedded withone or more coagulant and/or vasoconstrictor agent(s). In certainembodiments, a kit comprises pharmaceutical compositions comprising abarrier-forming material embedded or combined with one or more coagulantand/or vasoconstrictor agent(s), wherein the barrier-forming materialcontains an adhesive so that the material can be adhere to a patient'sskin surface. Alternatively, a kit lacks barrier-forming materials.

A kit can comprise a notice regarding FDA approval and/or instructionsfor use at a distance and/or in combination with compression.

A kit may be prescribed, for example, to patients requiringanticoagulant therapy, to avert the risk of serious bleeding which canoccur from minor injury. A kit may be used, for example, to treat abreach or puncture in a vein or artery resulting from catheterization.In one embodiment a kit comprises a patch embedded or coated with avasoconstrictor, a coagulant, or an agent that functions as both avasoconstrictor or coagulant. In another embodiment a kit comprises agel that can be used in the methods of the invention, wherein the gelcomprises a vasoconstrictor, a coagulant, or an agent that functions asboth a vasoconstrictor or coagulant.

Additionally, a kit designed for emergency or military use can alsocontain disposable pre-sterilized instruments, such as scissors,scalpel, clamp, tourniquet, elastic or inelastic bandages, or the like.In a preferred embodiment the kit contains gauze.

6. EXAMPLE 1 Determining Rate of Cessation or Time to Achieve Cessationof Blood Flow

This blinded, randomized, placebo controlled trial is designed toexamine an arterial puncture of a femoral artery in dogs intended tosimulate cardiac catheterization. This method takes into considerationdifferences in bleeding time and the amount of compression applied.

6.1 Materials and Methods

Dogs are randomly assigned to have either compression in conjunctionwith a topical barrier-forming material without a vasoconstrictor orcoagulant or a barrier-forming material coated with or formulated fromthe composition of the invention in conjunction with compression.

In preferred embodiments, the animal model provides a control. Forexample, in dogs the femoral arteries in each hind leg can be puncturedin an identical manner to create comparable punctures and the treatmentbeing tested can be applied to one puncture while the other is leftuntreated or receives compression alone.

A mechanical pressure clamp is applied over the barrier-formingmaterial. At 5 minute intervals, the clamp is loosened to check forblood flow from the wound site, but the barrier-forming material is notdisturbed. If blood continues to flow from the wound, the clamp isre-applied until bleeding completely stops, with checks performed everyfive minutes. A fluid-filled balloon connected to a pressure transduceris used to measure the amount of pressure applied. A pre-procedurebleeding time is also preformed.

By comparing pre-procedure bleeding variables such as pre-procedurehematocrit (%) and bleeding time (mins), for each group differences inbleeding can be eliminated through selection of particular animals, orcompensated for by calculating the rate of cessation or time to achievecessation of bleeding from a base values.

Clamp pressure applied to the femoral arterial puncture site can becarefully controlled and recorded for each dog, allowing calculation ofa mean clamp pressure (mmHg) for both the placebo group and the groupreceiving the composition of the invention.

The superficial nature of the femoral artery in dogs allows for directand consistent visualization of hematoma formation. Visualization can beby any of the methods described herein such as ultrasonography, orscintigraphic imaging. External signs are also indicative of hematomaformation, including ecchymosis (blue or purplish skin discolorationgreater than or equal to 2.5 cm in diameter), swelling (greater than orequal to 2.5 cm in diameter and 0.5 cm in height). The cannula can alsobe removed from the dogs following application of the methods of theinvention to determine frequency of hematomas or other vascularcomplications.

Percent differences between mean values for each treatment group arecalculated for several variables including, pre-procedure hematocrit(%), pre-procedure bleeding time (mins), time to cessation of bleedingfrom the puncture site (mins), the mean numbers of hematomas, and meanclamp pressure (mmHg). The statistical significance (P values) of thedifferences between the mean values for each variable are calculated bystandard statistical methods.

The present invention is not to be limited in scope by the specificembodiments described herein. Indeed, various modifications of theinvention in addition to those described will become apparent to thoseskilled in the art from the foregoing description and accompanyingfigures. Such modifications are intended to fall within the scope of theappended claims.

Various publications are cited herein, the disclosures of which areincorporated by reference in their entireties.

The present invention is not to be limited in scope by the specificembodiments described herein. Indeed, various modifications of theinvention in addition to those described will become apparent to thoseskilled in the art from the foregoing description and accompanyingfigures. Such modifications are intended to fall within the scope of theappended claims.

Various publications are cited herein, the disclosures of which areincorporated by reference in their entireties.

The invention can be illustrated by the following embodiments enumeratedin the numbered paragraphs that follow:

1. A method for treating a breach or puncture in a vein or artery of apatient, comprising:

-   -   a) applying topically to the patient's skin over a wound        contiguous with the breach or puncture in the vein or artery a        composition comprising an effective amount of a vasoconstrictor,        wherein the vasoconstrictor does not comprise a poly-β-1→4        N-acetylglucosamine polymer or derivative thereof; and        concurrently    -   b) applying compression to the breached or punctured vein or        artery,

wherein a cessation or reduction of blood flow out of the breach orpuncture in the vein or artery is achieved at a greater rate or in lesstime than applying compression in conjunction with a topicalbarrier-forming material without the vasoconstrictor.

2. A method for achieving a cessation of blood flow or sealing of abreach or puncture in a vein or artery and a cessation of blood flow orsealing of a skin surface wound that is contiguous with the breach orpuncture comprising:

-   -   a) applying topically to the patient's skin over a wound        contiguous with a breach or puncture in a vein or artery a        composition comprising a vasoconstrictor or coagulant, wherein        the vasoconstrictor or coagulant does not comprise a poly-β-1→4        N-acetylglucosamine polymer or derivative thereof;    -   b) concurrently applying compression to the breached or        punctured vein or artery; and    -   c) recording the amount of blood flow from the wound and the        puncture,

wherein an amount of the vasoconstrictor or coagulant is effective toincrease sealing or decrease cessation of blood flow from the breach orpuncture in the vein or artery and increase sealing or decreasecessation of blood flow from the skin surface wound, in comparison toapplying compression in conjunction with a topical barrier-formingmaterial without the vasoconstrictor.

3. A method for treating a breach or puncture in a vein or artery of apatient, comprising:

a) applying topically to the patient's skin over a wound contiguous withthe breach or puncture in the vein or artery a composition comprising aneffective amount of a coagulant, wherein the coagulant does not comprisea poly-β-1→4 N-acetylglucosamine polymer or derivative thereof; andconcurrently

b) applying compression to the breached or punctured vein or artery,

wherein a cessation or reduction of blood flow out of the breach orpuncture in the vein or artery is achieved at a greater rate in lesstime than applying compression in conjunction with a topicalbarrier-forming material without a coagulant.

4. A method for treating a breach or puncture in a vein or artery of apatient, comprising:

-   -   a) applying topically to the patient's skin over a wound        contiguous with the breach or puncture in the vein or artery a        composition comprising an effective amount of a vasoconstrictor        and a coagulant, wherein the vasoconstrictor and the coagulant        do not comprise a poly-β-1→4 N-acetylglucosamine polymer or        derivative thereof; and concurrently    -   b) applying compression to the breached or punctured vein or        artery,

wherein a cessation or reduction of blood flow out of the breach orpuncture in the vein or artery is achieved at a greater rate or in lesstime than applying compression in conjunction with a topicalbarrier-forming material without the vasoconstrictor and the coagulant.

5. The method of any one of paragraphs 1-4, wherein the compositionfurther comprises an anti-fungal or antibacterial agent.

6. The method of any one of paragraphs 1-4, wherein the compositionfurther comprises collagen.

7. The method of any one of paragraphs 1-4, wherein the compositionfurther comprises a pharmaceutical carrier.

8. The method of any one of paragraphs 1-4, wherein the composition isformulated as a gel, solid, liquid, sponge, foam, spray, emulsion,suspension, or solution.

9. The method of any one of paragraphs 1-4, wherein the compositionfurther comprises a neutral liquid, neutral gel or neutral solid.

10. The method of paragraph 9, wherein the composition further comprisesa neutral solid and wherein the neutral solid is a gauze.

11. The method of paragraph 8, wherein the composition is in the form ofa coating on a neutral solid.

12. The method of paragraph 11, wherein the neutral solid is a gauze.

13. The method of any one of paragraphs 1-4, wherein the barrier-formingmaterial is a gauze.

14. The method of any one of paragraphs 2, 3, or 4, wherein thecoagulant is selected from the group consisting of alpha-2-antiplasmin,alpha-1-antitrypsin, alpha-2-macroglobulin, aminohexanoic acid,aprotinin, a source of Calcium ions, calcium alginate, calcium-sodiumalginate, casein Kinase II, chitin, chitosan, collagen, cyanoacrylates,epsilon-aminocaproic acid, Factor XIII, fibrin, fibrin glue, fibrinogen,fibronectin, gelatin, living platelets, metha crylates, PAI-1, PAI-2,plasmin activator inhibitor, plasminogen, platelet agonists, protaminesulfate, prothrombin, an RGD peptide, sphingosine, a sphingosinederivative, thrombin, thromboplastin, and tranexamic acid.

15. The method of any one of paragraphs 1 or 4, wherein thevasoconstrictor is selected from the group consisting of adrenaline,endothelin-1, epinephrine, phenylephrine, serotonin, thromboxane, andU-46619.

16. The method of any one of paragraphs 1-4, wherein the patient is ahuman.

17. The method of any one of paragraphs 1-4, wherein composition appliedis a film or membrane.

18. The method of paragraph 17, wherein the film or membrane comprises abarrier-forming material.

19. The method of any one of paragraphs 1-4, wherein composition isformulated as a mat, string, microbead, microsphere, or microfibril.

20. The method of any one of paragraphs 1-4, wherein the compositionfurther comprises a biodegradable material.

21. The method of paragraph 20, wherein the biodegradable material isselected from the group consisting of a polyanionic polysaccharide,alginic acid, collagen, a polypeptide, a polyglycolide, a polylactide, apolycaprolactone, dextran and a copolymer of dextran, a polyglycolide, apolylactide, a polydioxanone, a polyestercarbonate, apolyhydroxyalkonate, and a polycaprolactone and a copolymer thereof.

22. The method of any one of paragraphs 1-4, further comprising beforestep (a) the step of administering to the patient an anticoagulant.

23. The method of paragraph 22, wherein the anticoagulant is selectedfrom the group consisting of coumadin, heparin, nadroparin, asparin, anda thrombolytic agent.

24. The method of paragraph 23, wherein the composition furthercomprises protamine sulfate in an amount effective to neutralizeheparin.

25. The method of any one of paragraphs 1-4, wherein the artery is thefemoral, radial, brachial, or axillary artery.

26. The method of any one of paragraphs 1-4, wherein the vein is thefemoral, internal jugular, or subclavian vein.

27. The method of any one of paragraphs 1-4, wherein the compression ismanual compression.

28. The method of any one of paragraphs 1-4, wherein the compression ismechanical compression.

29. The method of any one of paragraphs 1-4, wherein the compression isapplied to the vein or artery proximal of the puncture or breach.

30. The method of any one of paragraphs 1-4, wherein the compression isapplied at the site of application of the composition.

31. The method of any one of paragraphs 1-4, wherein the compression isapplied with a compression bandage.

32. The method of any one of paragraphs 1-4, further comprising,repeating step (b).

33. The method of paragraph 32, wherein the rate is at least 10% greaterthan applying compression in conjunction with a topical barrier-formingmaterial without a vasoconstrictor.

4. The method of paragraph 32, wherein the rate is at least 20% greaterthan applying compression in conjunction with a topical barrier-formingmaterial without a vasoconstrictor.

35. The method of paragraph 32, wherein the rate is at least 30% greaterthan applying compression in conjunction with a topical barrier-formingmaterial without a vasoconstrictor.

36. The method of paragraph 32, wherein the rate is at least 40% greaterthan applying compression in conjunction with a topical barrier-formingmaterial without a vasoconstrictor.

37. The method of paragraph 32, wherein the rate is at least 50% greaterthan applying compression in conjunction with a topical barrier-formingmaterial without a vasoconstrictor.

38. The method of any one of paragraphs 1-4, wherein the vein or arteryis breached or punctured by a catheter.

39. The method of any one of paragraphs 1-4, wherein the skin woundcontiguous with the breach or puncture in the vein or artery is 10, 9,8, 7, 6, 5, or 4 cm from the puncture in the vein or artery.

40. A method for decreasing the occurrence of localized vascularcomplications comprising:

-   -   a) applying topically to the patient's skin over a wound        contiguous with a breach or puncture in a vein or artery a        composition comprising a vasoconstrictor or coagulant, wherein        the vasoconstrictor or coagulant does not comprise a poly-β-1→4        N-acetylglucosamine polymer or derivative thereof;    -   b) concurrently applying compression to the breached or        punctured vein or artery; and    -   c) recording the occurrence of localized vascular complications,

wherein an amount of the vasoconstrictor or coagulant is effective tocause sealing of the breach or puncture in the vein or artery, reducingthe rate of localized vascular complications in comparison to applyingcompression in conjunction with a topical barrier-forming materialwithout a vasoconstrictor.

41. The method of paragraph 40, wherein the rate is 50% less thanapplying compression in conjunction with a topical barrier without avasoconstrictor.

42. The method of paragraph 40, wherein the vein or artery is breachedor punctured by a catheter.

43. A pharmaceutical composition for topically treating a breach orpuncture in a vein or artery, comprising:

-   -   a) an effective amount of a vasoconstrictor, wherein the        vasoconstrictor does not comprise a poly-β-1→4        N-acetylglucosamine polymer or derivative thereof; and    -   b) a pharmaceutically acceptable carrier.

44. The pharmaceutical composition of paragraph 43, wherein thecomposition further comprises a coagulant.

45. The pharmaceutical composition of paragraphs 43 or 44, furthercomprising a neutral liquid, neutral gel or neutral solid.

46. The pharmaceutical composition of paragraph 45, wherein thecomposition further comprises a neutral solid and wherein the neutralsolid is a gauze.

47. A kit comprising in one or more containers a composition comprising:

-   -   a) an effective amount of a vasoconstrictor, wherein the        vasoconstrictor does not comprise a poly-β-1→4        N-acetylglucosamine polymer or derivative thereof; and    -   b) a pharmaceutically acceptable carrier,        and instructions for topically treating a breach or puncture in        a vein or artery.

48. The kit of paragraph 47, wherein the composition further comprises acoagulant.

49. The kit of paragraph 47, further comprising a neutral liquid,neutral gel or neutral solid.

50. The kit of paragraph 49, wherein the composition further comprises aneutral solid and wherein the neutral solid is a gauze.

51. The kit of paragraph 49, wherein the composition further comprises aneutral gel and wherein the neutral gel is gelatin.

52. A pharmaceutical composition for topically treating a breach orpuncture in a vein or artery, comprising:

-   -   a) an effective amount of a vasoconstrictor, wherein the        vasoconstrictor does not comprise a poly-β-1→4        N-acetylglucosamine polymer or derivative thereof; and    -   b) wherein the vasoconstrictor is formulated into a barrier,        membrane, or film.

53. The pharmaceutical composition of paragraph 52, wherein thecomposition further comprises a coagulant.

54. The pharmaceutical composition of paragraphs 52 or 53, furthercomprising a neutral liquid, neutral gel or neutral solid.

55. The pharmaceutical composition of paragraph 52, wherein thecomposition further comprises a neutral solid and wherein the neutralsolid is a gauze.

56. A kit comprising in one or more containers a composition comprising:

-   -   a) an effective amount of a vasoconstrictor, wherein the        vasoconstrictor does not comprise a poly-β-1→4        N-acetylglucosamine polymer or derivative thereof; and    -   b) wherein the vasoconstrictor is formulated into a barrier,        membrane, or film,

and instructions for topically treating a breach or puncture in a veinor artery.

57. The kit of paragraph 56, wherein the composition further comprises acoagulant.

58. The kit of paragraph 56, wherein the composition further comprises aneutral liquid, neutral gel or neutral solid.

59. The kit of paragraph 56, wherein the composition further comprises aneutral solid and wherein the neutral solid is a gauze.

60. The kit of paragraph 56, wherein the vasoconstrictor is Adrenaline™and the composition comprises about 0.00001 mg/kg to about 0.5 mg/kg ofpatient body weight of Adrenaline™.

61. The kit of paragraph 56, wherein the vasoconstrictor is metaraminolbitartrate and the composition comprises about 0.0005 mg/kg to about 4.5mg/kg of patient body weight of metaraminol bitartrate.

62. The kit of paragraph 56, wherein the vasoconstrictor is dopamine HCland the composition comprises about 0.0005 mg/kg to about 10 mg/kg ofpatient body weight of dopamine HCl.

63. The kit of paragraph 56, wherein the vasoconstrictor isisoproterenol HCl and the composition comprises about 0.0005 mg/kg toabout 5 mg/kg of patient body weight of isoproterenol HCl.

64. The kit of paragraph 56, wherein the vasoconstrictor isnorepinephrine and the composition comprises about 0.0001 mg/kg to about0.01 mg/kg of norepinephrine.

65. The kit of paragraph 56, wherein the vasoconstrictor is Serotonin™and the composition comprises about 0.001 mg/kg to about 0.6 mg/kg ofpatient body weight of Serotonin™.

66. The kit of paragraph 56, wherein the vasoconstrictor is endothelinand the composition comprises about 4.5×10⁻⁸ mg/kg to about 5.0×10⁻⁶mg/kg of patient body weight of endothelin.

What is claimed is:
 1. A method for inhibiting the formation ofhematomas resulting from a cardiac catheterization procedure in apatient, comprising: a) applying topically over a catheter exit site onthe skin of a patient in need of such inhibition a compositioncomprising an effective amount of one or more vasoconstrictors, whereinthe one or more vasoconstrictors do not comprise a poly-β-1→4N-acetylglucosamine polymer or derivative thereof, and wherein thecatheter exit site is contiguous with a puncture in a vein or arterymade by the catheter by 1-10 cm; and concurrently b) applyingcompression to the punctured vein or artery.
 2. The method of claim 1,wherein the vasoconstrictor is endothelin, endothelin-1, epinephrine,adrenaline, metaraminol bitartrate, dopamine HCl, isoproterenol HCl,norepinephrine, phenylephrine, serotonin, thromboxane, prostaglandin,methergine, oxytocin, isopreland U-46619, papaverine, yohimbine,visnadin, khellin, bebellin, or a nicotinate derivative.
 3. The methodof claim 1, wherein the composition further comprises an anti-fungal orantibacterial agent.
 4. The method of claim 1, wherein the compositionfurther comprises collagen.
 5. The method of claim 1, wherein thecomposition further comprises a pharmaceutical carrier.
 6. The method ofclaim 1, wherein the composition is formulated as a gel, solid, liquid,sponge, foam, spray, emulsion, suspension, or solution.
 7. The method ofclaim 6, wherein the composition is in the form of a coating on aneutral solid.
 8. The method of claim 7, wherein the neutral solid is agauze.
 9. The method of claim 1, wherein the composition furthercomprises a neutral liquid, neutral gel or neutral solid.
 10. The methodof claim 9, wherein the composition further comprises a neutral solidand wherein the neutral solid is a gauze.
 11. The method of claim 1,wherein the composition is formulated as a barrier-forming material. 12.The method of claim 1, wherein the composition further comprises one ormore coagulants.
 13. The method of claim 12, wherein the one or morecoagulants are alpha-2-antiplasmin, alpha-1-antitrypsin,alpha-2-macroglobulin, aminohexanoic acid, aprotinin, a source ofCalcium ions, calcium alginate, calcium-sodium alginate, casein KinaseII, chitin, chitosan, collagen, cyanoacrylates, epsilon-aminocaproicacid, Factor XIII, fibrin, fibrin glue, fibrinogen, fibronectin,gelatin, living platelets, metha crylates, PAI-1, PAI-2, plasminactivator inhibitor, plasminogen, platelet agonists, protamine sulfate,prothrombin, an RGD peptide, sphingosine, a sphingosine derivative,thrombin, thromboplastin, or tranexamic acid.
 14. The method of claim 1,wherein the patient is a human.
 15. The method of claim 1, wherein thecomposition comprises a film or membrane.
 16. The method of claim 15,wherein the film or membrane comprises a barrier-forming material. 17.The method of claim 1, wherein composition is formulated as a mat, astring, a microbead, a microsphere, or a microfibril.
 18. The method ofclaim 1, wherein the composition further comprises one or morebiodegradable materials.
 19. The method of claim 18, wherein the one ormore biodegradable materials are a polyanionic polysaccharide, alginicacid, collagen, a polypeptide, a polyglycolide, a polylactide, apolycaprolactone, dextran, a copolymer of dextran, a polyglycolide, apolylactide, a polydioxanone, a polyestercarbonate, apolyhydroxyalkonate, a polycaprolactone, or a copolymer thereof.
 20. Themethod of claim 1, further comprising before step (a) the step ofadministering to the patient an anticoagulant.
 21. The method of claim20, wherein the anticoagulant comprises one or more of coumadin,heparin, nadroparin, aspirin, or a thrombolytic agent.
 22. The method ofclaim 21, wherein the composition further comprises protamine sulfate inan amount effective to neutralize heparin.
 23. The method of claim 1,wherein the artery is the femoral, radial, brachial, or axillary artery.24. The method of claim 1, wherein the vein is the femoral, internaljugular, or subclavian vein.
 25. The method of claim 1, wherein thecompression is manual compression.
 26. The method of claim 1, whereinthe compression is mechanical compression.
 27. The method of claim 1,wherein the compression is applied to the vein or artery proximal of thepuncture in the vein or artery.
 28. The method of claim 1, wherein thecompression is applied at the site of application of the composition.29. The method of claim 1, wherein the compression is applied with acompression bandage.
 30. The method of claim 1, further comprisingrepeating step (b).
 31. The method of claim 1, wherein the catheter exitsite on the skin contiguous with the puncture in the vein or artery is10, 9, 8, 7, 6, 5, or 4 cm from the puncture in the vein or artery. 32.The method of claim 1, wherein the compression is applied for at leastone time interval of up to ten minutes.
 33. The method of claim 1,wherein the vasoconstrictor is endothelin, endothelin-1, metaraminolbitartrate, dopamine HCl, isoproterenol HCl, norepinephrine,phenylephrine, serotonin, thromboxane, prostaglandin, methergine,oxytocin, isopreland U-46619, papaverine, yohimbine, visnadin, khellin,bebellin, or a nicotinate derivative.
 34. A method for decreasing theoccurrence of localized vascular complications comprising: a) applyingtopically over a wound on the skin of a patient at risk of saidcomplications a composition comprising an effective amount of one ormore vasoconstrictors, wherein the one or more vasoconstrictors do notcomprise a poly-β-1→4 N-acetylglucosamine polymer or derivative thereof,and wherein the wound is contiguous with a breach or puncture in a veinor artery; and concurrently b) applying compression to the breached orpunctured vein or artery.
 35. The method of claim 34, wherein the veinor artery is breached or punctured by a catheter.
 36. The method ofclaim 35, wherein the breach or puncture results from a cardiaccatheterization procedure.
 37. The method of claim 35, wherein the woundon the skin is contiguous with the breach or puncture made by thecatheter by 1-10 cm.
 38. The method of claim 36, wherein thevasoconstrictor is endothelin, endothelin-1, epinephrine, adrenaline,metaraminol bitartrate, dopamine HCl, isoproterenol HCl, norepinephrine,phenylephrine, serotonin, thromboxane, prostaglandin, methergine,oxytocin, isopreland U-46619, papaverine, yohimbine, visnadin, khellin,bebellin, or a nicotinate derivative.
 39. The method of claim 34,wherein the compression is applied for at least one time interval of upto ten minutes.
 40. The method of claim 34, wherein the wound on theskin is contiguous with the breach or puncture in the vein or artery by1-10 cm.
 41. The method of claim 34, wherein the vasoconstrictor isendothelin, endothelin-1, epinephrine, adrenaline, metaraminolbitartrate, dopamine HCl, isoproterenol HCl, norepinephrine,phenylephrine, serotonin, thromboxane, prostaglandin, methergine,oxytocin, isopreland U-46619, papaverine, yohimbine, visnadin, khellin,bebellin, or a nicotinate derivative.
 42. The method of claim 34,wherein the composition further comprises one or more coagulants. 43.The method of claim 34, further comprising before step (a) the step ofadministering to the patient an anticoagulant.